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元件指南
VectorBuilder提供了许多常用的载体元件，可供用户在设计载体时进行选择。下表按类型提供了这些常用元件的详细信息。
Stuffer
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| ORF_Stuffer | Amino acid 2-83 of E. coli beta-galactosidase | Can be used as a negative ORF control. | Designed by VectorBuilder | View |
荧光报告基因
| 姓名 | 描述 | Application Notes | Color | Structure | Maximum Excitation (nm) | Maximum Emission (nm) | Brightness (% of EGFP) | References | 序列 |
|---|---|---|---|---|---|---|---|---|---|
| EGFP | Enhanced green fluorescent protein; codon optimized based on a variant of wild type GFP from the jellyfish Aequorea victoria | Commonly used green fluorescent protein; ranked high in brightness, photostability and pH stability among all fluorescent proteins. | Green | Monomer (may form weak dimer) | 484 | 507 | 100 | Nucleic Acids Res. 24:4592 (1996) | View |
| NLS-EGFP | EGFP with nuclear localization signal on both ends | Nuclear localization. | Green | Monomer (may form weak dimer) | 484 | 507 | 100 | Engineered by VectorBuilder | View |
| sfGFP | Superfolder green fluorescent protein from Aequorea victoria; GFP variant S30R, Y39N, N105T, Y145F, I171V and A206V | Folds well when fused with a variety of other polypeptides; fast folding, highly stable. | Green | Monomer (may form weak dimer) | 485 | 510 | 160 | Nat Biotechnol. 24:79 (2006) | View |
| EmGFP | Emerald green fluorescent protein; variant of EGFP generated by mutagenesis | Enhanced photostability and brightness compared to its predecessor EGFP. | Green | Monomer (may form weak dimer) | 487 | 509 | 116 | Methods Cell Biol. 58:19 (1999) | View |
| TurboGFP | Also known as maxGFP; green fluorescent protein from maxillopoda | Fast maturation; ranked high in brightness, photostability and pH stability among all fluorescent proteins. | Green | Dimer | 482 | 502 | 112 | Mol Biol Evol. 21:841 (2004) | View |
| hrGFP | Humanized recombinant green fluorescent protein derived from Renilla reniformis | Low cytotoxicity. | Green | Monomer | 500 | 506 | 100 | J Biol Chem. 254:781 (1979) | View |
| d2EGFP | Destabilized EGFP due to addition of PEST sequence from mouse Odc1 gene | Fast turnover (fluorescence half life is 2 hours). | Green | Monomer | 488 | 507 | 100 | J Biol Chem. 273:34970 (1998) | View |
| ZsGreen1 | Bright green fluorescent protein derived from a Zoanthus sp. reef coral | High solubility; bright emission; rapid chromophore maturation. | Green | Tetramer | 493 | 505 | 250 | Nat Biotechnol. 17:969 (1999); J Histochem Cytochem. 55:931 (2007) | View |
| EGFP(S65T) | Enhanced green fluorescent protein with S65T mutation | Higher expression in plant than GFP | Green | Monomer (may form weak dimer) | 484 | 507 | 100 | Curr Biol. 6:325 (1996) | View |
| mNeonGreen | Yellow-green fluorescent protein derived from a tetrameric fluorescent protein from the cephalochordate Branchiostoma lanceolatum | Brightest monomeric green or yellow fluorescent protein; superior photostability; good acid tolerance; fast maturation; excellent fusion tag for traditional imaging as well as stochastic single-molecule super resolution imaging; can be used for FRET. | Green or Yellow | Monomer | 506 | 517 | 276 | Nat Methods. 10:407 (2013) | View |
| Venus | Variant of yellow fluorescent protein (YFP) generated by mutagenesis; YFP is itself a variant of EGFP generated by mutagenesis | Fast maturation and high tolerance to acidosis and Cl- compared to its predecessor YFP. | Yellow | Monomer (may form weak dimer) | 515 | 528 | 156 | Nat biotechnol. 20:87 (2002) | View |
| EYFP | Enhanced yellow fluorescent protein, GFP variant S65G/V68LS72A/T203Y | Excitaton and emission are sensitive to pH; can be pH indicator in cytosol and nucleus. | Yellow | Monomer (may form weak dimer) | 513 | 527 | 150 | Science. 273: 1392 (1996); Structure. 6:1267 (1998) | View |
| YPet | Derived by combining mutations from Venus and YFP3; all these variants trace their origins to EGFP | Reduced pH sensitivity but slower folding rate compared to its predecessor Venus; can be used in conjunction with CyPet for FRET applications. | Yellow | Monomer (may form weak dimer) | 517 | 530 | 238 | Nat Biotechnol. 23:355 (2005) | View |
| Cerulean | Improved ECFP | With improved signal-to-noise ratio; 2.5-fold brighter than ECFP; can be used for FRET. | Cyan | Monomer (may form weak dimer) | 433 | 475 | 79 | Nat Biotechnol. 22:445 (2004) | View |
| CyPet | Variant of cyan fluorescent protein (CFP) generated by mutagenesis; CFP is itself a variant of EGFP generated by mutagenesis | Improved protein folding rate compared to its predecessor CFP; can be used in conjunction with Ypet for FRET applications. | Cyan | Monomer (may form weak dimer) | 435 | 477 | 53 | Nat Biotechnol. 23:355 (2005) | View |
| AmCyan | Cyan fluorescent protein variant derived from Anemonia majano with N34S and K68M mutations | Brighter than ECFP. | Cyan | Tetramer | 458 | 489 | 31 | Nat Biotechnol. 17:969 (1999) | View |
| EBFP | Blue variant of EGFP generated by mutagenesis | Low fluorescence and low photostability. | Blue | Monomer (may form weak dimer) | 383 | 445 | 27 | Biochem. 46:5904 (2007) | View |
| TagBFP | Also known as mTagBFP; blue variant of TagRFP generated by mutagenesis | Rank high in brightness, photostability and pH stability among blue fluorescent proteins. | Blue | Monomer (may form weak dimer) | 402 | 457 | 99 | Chem Biol. 15:1116 (2008) | View |
| TagBFP2 | Also known as mTagBFP2; improved variant of TagBFP generated by I174A mutation | Higher stability and brightness; rapid chromophore formation. | Blue | Monomer (may form weak dimer) | 399 | 454 | 121 | PLoS One. 6: e28674 (2011) | Ver 1 Ver 2 Ver 3 |
| NLS-TagBFP2 | TagBFP2 with nuclear localization signals on both ends | Nuclear localization. | Blue | Monomer (may form weak dimer) | 399 | 454 | 121 | Engineered by VectorBuilder | View |
| dTomato | Dimeric variant of the Discosoma sp. red fluorescent protein (DsRed) generated by mutagenesis | Fast maturation but slightly lower brightness compared to its predecessor DsRed. | Orange | Dimer | 554 | 581 | 142 | Nat Biotechnol. 22:1567 (2004) | View |
| tdTomato | Tandem dTomato generated by the fusion of two copies dTomato. | Exceptionally bright red fluorescent protein; low aggregation and low acid sensitivity. | Orange | Monomer (nonaggregating tandem dimer) | 554 | 581 | 283 | Nat Biotechnol. 22:1567 (2004) | View |
| NLS-tdTomato | tdTomato with nuclear localization signals on both ends | Nuclear localization. | Orange | Monomer (nonaggregating tandem dimer) | 554 | 581 | 283 | Engineered by VectorBuilder | View |
| DsRed_Express2 | Fast maturing variant of the Discosoma sp. red fluorescent protein (DsRed) generated by mutagenesis | High solubility, fast maturation, reduced green emission and reduced cytotoxicity compared to its predecessor DsRed. | Orange | Tetramer | 554 | 591 | 72 | Nat Biotechnol. 20:83 (2002) | View |
| TurboRFP | Derived from the sea anemone Entacmaea quadricolor | Fast maturation; ranked high in brightness and pH stability among red fluorescent proteins. | Orange | Dimer | 553 | 574 | 187 | Nat Methods. 4:555 (2007) | View |
| mRFP1 | Monomeric red fluorescent protein 1; variant of DsRed generated by mutagenesis | Fast maturation but lower brightness compared to its predecessor DsRed. | Red | Monomer | 584 | 607 | 37 | Proc Natl Acad Sci U S A. 99:7877 (2002) | View |
| mCherry | Variant of mRFP1 generated by mutagenesis | Commonly used red fluorescent protein; fast maturation compared to its predecessor, mRFP1. | Red | Monomer | 587 | 610 | 47 | Nat Biotechnol. 22:1567 (2004) | Ver 1 Ver 2 |
| NLS-mCherry | mCherry with nuclear localization signals on both ends | Nuclear localization. | Red | Monomer | 587 | 610 | 47 | Engineered by VectorBuilder | View |
| mApple | Photostable monomeric derivative of DsRed | Brighter than mCherry; constant photostability. | Red | Monomer | 568 | 592 | 109 | Nat Methods. 5:545 (2008) | View |
| mKate2 | Katushka2 far-red fluorescent protein | Fast maturation; high pH-stability and photostability; low cytotoxicity; a superior fluorescent tag for imaging in living tissues. | Red | Monomer | 588 | 633 | 74 | Biochem J. 418: 567 (2009) | View |
| EGFP+IVS | EGFP inserted with 3 synthetic introns | Insertion of additional intron sequences results in increased gene expression in C. elegans. | Green | Monomer (may form weak dimer) | 484 | 507 | 100 | Curr Biol. 18:1476 (2008) | View |
| EGFP/H2B | EGFP fused to histone H2B | Allows to be localized efficiently to chromatin and facilitates detection. | Green | Monomer (may form weak dimer) | 484 | 507 | 100 | Curr Biol. 18:1476 (2008) | View |
| mCherry+IVS | mCherry inserted with 3 synthetic introns | Insertion of additional intron sequences results in increased gene expression in C. elegans. | Red | Monomer | 587 | 610 | 47 | Curr Biol. 18:1476 (2008) | View |
| mCherry/H2B | mCherry fused to histone H2B | Allows to be localized efficiently to chromatin and facilitates detection. | Red | Monomer | 587 | 610 | 47 | Curr Biol. 18:1476 (2008) | View |
| miRFP670nano | Near-infrared fluorescent protein evolved from cyanobacteriochrome photoreceptor NpR3784 | Small protein exhibited high binding efficiency of endogenous biliverdin chromophore; highly stable to denaturation and degradation and can be used as an internal protein tag; can be used for FRET. | Red | 645 | 670 | / | Nat Commun. 10:279 (2019) | View |
抗药性基因
| 姓名 | 描述 | Selection Drug | Usage | Application Notes | References | 序列 |
|---|---|---|---|---|---|---|
| Neo | Neomycin resistance gene | Geneticin (G418) | Positive selection | Allows cells to be resistant to geneticin (G418). | Proc Natl Acad Sci U S A. 81:6466 (1984) | View |
| Puro | Puromycin resistance gene | Puromycin | Positive selection | Allows cells to be resistant to puromycin. | Gene. 62:121 (1988) | View |
| Hygro | Hygromycin resistance gene | Hygromycin B | Positive selection | Allows cells to be resistant to hygromycin B. | Gene. 56:117 (1987) | Ver 1 Ver 2 Ver 3 |
| Bsd | Blasticidin resistance gene | Blasticidin | Positive selection | Allows cells to be resistant to blasticidin. | Biochim Biophys Acta. 1219:653 (1994) | Ver 1 Ver 2 |
| Bleo | Bleomycin resistance gene | Bleomycin, phleomycin or Zeocin(TM) | Positive selection | Allows cells to be resistant to bleomycin, phleomycin or Zeocin(TM). | Somat Cell Mol Genet. 14:243 (1988) | View |
| Bar | Phosphinothricin acetyltransferase gene | Glufosinate (phosphinothricin) | Positive selection | Allows plant to be resistant to phosphinothricin. | Plant Mol Biol. 21:871 (1993) | View |
| Neo/Kana | Neomycin phosphotransferase II gene | G418 or Kanamycin | Positive selection | Allows plant to be resistant to G418 or Kanamycin. | Methods Mol Biol. 44:201 (1995) | View |
| Hygro | Hygromycin resistance gene | Hygromycin B | Positive selection | Allows plant to be resistant to Hygromycin B. | Biotechnol Lett. 29:1793 (2007) | View |
注意
:
- Some components have different versions that differ slightly in sequence but are functionally equivalent. VectorBuilder may use different versions depending on context.
自杀基因
| 姓名 | 描述 | Inducer | Application Notes | References | 序列 |
|---|---|---|---|---|---|
| FKBP/Casp8 | FK506-binding protein 12 (with F36V mutation) fused with human caspase-8 protein deleted for 1-215 AA and linked to the N-myristoylation signal from Src kinase | Dimerization drug, e.g. AP20187 | Promotes rapid caspase-8 mediated apoptotic cell death upon AP20187 induced dimerization of FKBP-12. | J Biol Chem. 285: 16632 (2010) | View |
| iCasp9 | FK506-binding protein 12 (with F36V mutation) fused with human caspase-9 protein deleted for 1-134 AA and linked to a HA tag | Dimerization drug, e.g. AP20187 | Promotes rapid caspase-9 mediated apoptotic cell death upon AP20187 induced dimerization of FKBP-12. | Blood. 105:4247 (2005); N Engl J Med. 365:1673 (2011); Mol Ther. 26:1266 (2018) | View |
| deltaTK | Truncated thymidine kinase that converts ganciclovir (GCV) into a toxic metabolite that inhibits DNA synthesis | Ganciclovir (GCV) | Allows cells to be killed by ganciclovir (GCV); can be used in embryonic stem cells to generate knockout mice without causing sterility in mice. | Genesis. 28:31 (2000) | View |
| CodA | E.coli cytosine deaminase that catalyzes 5-fluorocytosine into toxic 5-fluorouracil | 5-fluorocytosine (5FC) | Allows cells to be killed by 5-fluorocytosine (5FC) | Proc Natl Acad Sci U S A. 89:33 (1992) | View |
| DTA | Diphtheria toxin A | - | Induces cell apoptosis by inhibiting EF-2 synthesis. | Proc Natl Acad Sci U S A. 87:9918 (1990) | View |
| DTR | Simian diphtheria toxin receptor | Diphtheria toxin (DT) | Cells expressing DTR are sensitive to diphtheria toxin (DT); DTR can be used for conditional and targeted cell ablation. | Cell. 69:1051 (1992) | View |
| peel-1 | C. elegans peel-1 gene | - | Ectopic expression of peel-1 at later life stages of C. elegans causes cell death and lethality. | PLoS Biol. 9:e1001115 (2011); Nat Methods. 9:117 (2012) | View |
双报告基因
| 姓名 | 描述 | 荧光 | Selection Drug | Application Notes | References | 序列 |
|---|---|---|---|---|---|---|
| EGFP/Neo | EGFP fused with Neo | Green | Geneticin (G418) | Allows cells to be visualized by green fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| EGFP/Puro | EGFP fused with Puro | Green | Puromycin | Allows cells to be visualized by green fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| EGFP/Hygro | EGFP fused with Hygro | Green | Hygromycin B | Allows cells to be visualized by green fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| EGFP/Bsd | EGFP fused with Bsd | Green | Blasticidin | Allows cells to be visualized by green fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| EGFP/Bleo | EGFP fused with Bleo | Green | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
| mCherry/Neo | mCherry fused with Neo | Red | Geneticin (G418) | Allows cells to be visualized by red fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | Ver 1 Ver 2 |
| mCherry/Puro | mCherry fused with Puro | Red | Puromycin | Allows cells to be visualized by red fluorescence and resistant to puromycin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| mCherry/Hygro | mCherry fused with Hygro | Red | Hygromycin B | Allows cells to be visualized by red fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| mCherry/Bsd | mCherry fused with Bsd | Red | Blasticidin | Allows cells to be visualized by red fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| mCherry/Bleo | mCherry fused with Bleo | Red | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
| TagBFP2/Neo | TagBFP2 fused with Neo | Blue | Geneticin (G418) | Allows cells to be visualized by blue fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2/Puro | TagBFP2 fused with Puro | Blue | Puromycin | Allows cells to be visualized by blue fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| TagBFP2/Hygro | TagBFP2 fused with Hygro | Blue | Hygromycin B | Allows cells to be visualized by blue fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| TagBFP2/Bsd | TagBFP2 fused with Bsd | Blue | Blasticidin | Allows cells to be visualized by blue fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2/Bleo | TagBFP2 fused with Bleo | Blue | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
| EGFP:T2A:Neo | EGFP and Neo linked by T2A | Green | Geneticin (G418) | Allows cells to be visualized by green fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| EGFP:T2A:Puro | EGFP and Puro linked by T2A | Green | Puromycin | Allows cells to be visualized by green fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| EGFP:T2A:Hygro | EGFP and Hygro linked by T2A | Green | Hygromycin B | Allows cells to be visualized by green fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| EGFP:T2A:Bsd | EGFP and Bsd linked by T2A | Green | Blasticidin | Allows cells to be visualized by green fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| EGFP:T2A:Bleo | EGFP and Bleo linked by T2A | Green | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | Ver 1 Ver 2 |
| mCherry:T2A:Neo | mCherry and Neo linked by T2A | Red | Geneticin (G418) | Allows cells to be visualized by red fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| mCherry:T2A:Puro | mCherry and Puro linked by T2A | Red | Puromycin | Allows cells to be visualized by red fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| mCherry:T2A:Hygro | mCherry and Hygro linked by T2A | Red | Hygromycin B | Allows cells to be visualized by red fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| mCherry:T2A:Bsd | mCherry and Bsd linked by T2A | Red | Blasticidin | Allows cells to be visualized by red fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| mCherry:T2A:Bleo | mCherry and Bleo linked by T2A | Red | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | Ver 1 Ver 2 |
| EGFP:P2A:Neo | EGFP and Neo linked by P2A | Green | Geneticin (G418) | Allows cells to be visualized by green fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| EGFP:P2A:Puro | EGFP and Puro linked by P2A | Green | Puromycin | Allows cells to be visualized by green fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| EGFP:P2A:Hygro | EGFP and Hygro linked by P2A | Green | Hygromycin B | Allows cells to be visualized by green fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| EGFP:P2A:Bsd | EGFP and Bsd linked by P2A | Green | Blasticidin | Allows cells to be visualized by green fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| EGFP:P2A:Bleo | EGFP and Bleo linked by P2A | Green | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
| mCherry:P2A:Neo | mCherry and Neo linked by P2A | mCherry | Geneticin (G418) | Allows cells to be visualized by mCherry fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| mCherry:P2A:Puro | mCherry and Puro linked by P2A | mCherry | Puromycin | Allows cells to be visualized by mCherry fluorescence and resistant to puromycin. | Engineered by VectorBuilder | View |
| mCherry:P2A:Hygro | mCherry and Hygro linked by P2A | mCherry | Hygromycin B | Allows cells to be visualized by mCherry fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | View |
| mCherry:P2A:Bsd | mCherry and Bsd linked by P2A | mCherry | Blasticidin | Allows cells to be visualized by mCherry fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| mCherry:P2A:Bleo | mCherry and Bleo linked by P2A | Red | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
| TagBFP2:T2A:Neo | TagBFP2 and Neo linked by T2A | Blue | Geneticin (G418) | Allows cells to be visualized by blue fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| TagBFP2:T2A:Puro | TagBFP2 and Puro linked by T2A | Blue | Puromycin | Allows cells to be visualized by blue fluorescence and resistant to puromycin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2:T2A:Hygro | TagBFP2 and Hygro linked by T2A | Blue | Hygromycin B | Allows cells to be visualized by blue fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2:T2A:Bsd | TagBFP2 and Bsd linked by T2A | Blue | Blasticidin | Allows cells to be visualized by blue fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| TagBFP2:T2A:Bleo | TagBFP2 and Bleo linked by T2A | Blue | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2:P2A:Neo | TagBFP2 and Neo linked by P2A | Blue | Geneticin (G418) | Allows cells to be visualized by blue fluorescence and resistant to geneticin (G418). | Engineered by VectorBuilder | View |
| TagBFP2:P2A:Puro | TagBFP2 and Puro linked by P2A | Blue | Puromycin | Allows cells to be visualized by blue fluorescence and resistant to puromycin. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2:P2A:Hygro | TagBFP2 and Hygro linked by P2A | Blue | Hygromycin B | Allows cells to be visualized by blue fluorescence and resistant to hygromycin B. | Engineered by VectorBuilder | Ver 1 Ver 2 |
| TagBFP2:P2A:Bsd | TagBFP2 and Bsd linked by P2A | Blue | Blasticidin | Allows cells to be visualized by blue fluorescence and resistant to blasticidin. | Engineered by VectorBuilder | View |
| TagBFP2:P2A:Bleo | TagBFP2 and Bleo linked by P2A | Blue | Bleomycin, phleomycin or Zeocin(TM) | Allows cells to be visualized by green fluorescence and resistant to Bleomycin, phleomycin or Zeocin(TM). | Engineered by VectorBuilder | View |
化学发光报告基因
| 姓名 | 描述 | Application Notes | Maximum Emission (nm) | References | 序列 |
|---|---|---|---|---|---|
| Luciferase | Firefly luciferase | Most commonly used luciferase. | 560 | Anal Biochem. 175:5 (1988) | View |
| Luc2 | Humanized firefly luciferase | Codon optimized to reduce cryptic transcription factor binding sites. | 560 | Promega Notes. 89:7 (2005) | View |
| MetLuc | Metridia luciferase | Secreted luciferase that allows detection in cell culture medium without lysing cells. | 480 | J Biol Chem. 279:3212 (2004) | View |
| Rluc | Renilla luciferase | Can be used as internal control in dual luciferase assay when paired with firefly luciferase. | 480 | Proc Natl Acad Sci U S A. 88:4438 (1991) | View |
| Nluc | Nano Luciferase | Small and super bright; can be used in dual luciferase assay when paired with firefly luciferase. | 460 | ACS Chem Biol. 7:1848 (2012) | View |
| Aequorin | Calcium-activated photoprotein isolated from the hydromedusan Aequorea victoria | Oxidizes coelenterazine (a luciferin molecule) into coelenteramide in the presence of Ca2+, emitting blue light in the process. | 465 | Proc Natl Acad Sci U S A. 82:3154 (1985) | View |
| hRluc | Humanized renilla luciferase | Can be used as internal control in dual luciferase assay when paired with firefly luciferase. | 480 | Biochemistry. 16:85 (1977); J Biomed Opt. 10:41210 (2005) | View |
| hRluc/Puro | Humanized renilla luciferase fused with puromycin resistance gene | Can be used as internal control in dual luciferase assay when paired with firefly luciferase and allows cells to be resistant to puromycin. | 480 | Engineered by VectorBuilder | View |
注意
:
- Luc2, Rluc and Nluc are trademarks of Promega.
显色报告基因
| 姓名 | 描述 | Application Notes | Substrate | Color | References | 序列 |
|---|---|---|---|---|---|---|
| LacZ | E. coli beta-galactosidase | High sensitivity. | X-gal or ONPG | Blue (X-gal); yellow (ONPG) | Microbiol Rev. 49:398 (1985) | Ver 1 Ver 2 |
| SEAP | Human secreted embryonic alkaline phosphatase | Secreted protein that allows detection in cell culture medium without lysing cells. | NBT/BCIP | Purple-blue | Gene. 66:1 (1988) | View |
| GUSPlus | A synthetic gusA gene based on the sequence from Staphylococcus sp., with a catalase intron inserted into the coding sequence to prevent expression in bacterial cells | A reporter to investigate gene transfer from Agrobacterium to plants; produces a GUS protein detectable in recipient plant. | X-Gluc | Blue | Nature. 433:629 (2005) | View |
注意
:
- Some components have different versions that differ slightly in sequence but are functionally equivalent. VectorBuilder may use different versions depending on context.
- GUSPlus contains an intron so that its length is not multiple of 3.
pH敏感型荧光报告基因
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| pHluorin2 | Enhanced, ratiometric, pH-sensitive green florescent protein | Has a bimodal excitation spectrum with peaks at 395 and 475 nm and an emission maximum at 509 nm; upon acidification, it has a dose-dependent decrease in the excitation at 395 nm with a corresponding increase in the excitation at 475 nm; it has a reversible excitation ratio change between pH 5.4~8.4. | Nature. 394:192 (1998); Adv Biosci Biotechnol. 2:132 (2011); Front Plant Sci. 4:523 (2013) | View |
| Superecliptic-pHluorin | Red-shifted ecliptic pHluorin that is excited strongly by blue light at neutral pH but minimal fluorescence at acidic pH | Displays a reversible excitation ratio change between pH 6.5~8.0. | Biophys J. 79:2199 (2000); Front Plant Sci. 4:523 (2013) | View |
| mt-mKeima | Monomeric dual-excitation Keima protein derived from stony coral Montipora by numerous cycles of semi-random mutagenesis; fused with a tandem repeat of mitochondria-targeting sequences from COX VIII | Localizes to the mitochondrial matrix for the detection of mitophagy using reporter color change. The excitation wavelength of the mKeima fluorescent protein is determined by the pH of the environment, with an excitation wavelength centered at 440 nm at neutral pH and an excitation of 586 nm at an acidic pH. The emission wavelength of Keima is identical at either pH and peaks maximally at 620 nm. | Chem Biol.18:1042 (2011); Nat Protoc.12:1576 (2017) | View |
转录调控蛋白
| 姓名 | 描述 | Application Notes | Target Sites | References | 序列 |
|---|---|---|---|---|---|
| GAL4 | Gal4 can bind to the galactose upstream activating sequence (UAS) to activate downstream genes expression | Gal4 can bind to the promoters which bear the galactose upstream activating sequence (UAS), then activate downstream genes expression. | UAS | Development. 118:401 (1993) | View |
| Gal4/VP16 | Yeast transcription activator protein Gal4 DNA binding domain fused to VP16 transactivation domain | Gal4 can bind to UAS to activate downstream gene transcription; this fusion protein of Gal4 and VP16 can strongly activate gene expression in zebrafish. | UAS | Dev Biol. 233:329 (2001) | View |
| VP16 | VP16 transcriptional activator domain encoded by HSV | Activates gene expression, commonly used with other DNA binding proteins. | Cell. 149:1447 (2012) | View | |
| VP64 | Tetrameric repeat of the minimal activation domain of VP16 | Strongly activates gene expression, commonly used with other DNA binding proteins. | Proc Natl Acad Sci U S A. 95: 14628 (1998); Proc Natl Acad Sci U S A. 97: 1495 (2000) | View | |
| scFv(GCN4)-sfGFP-GB1-VP64 | Anti-GCN4 single chain variable fragment (scFv) antibody fused to superfolder-GFP (sfGFP) with a GB1 solubility tag and VP64 transcriptional activator domain | Encodes a fusion protein consisting of GCN4 antibody, sfGFP and VP64; tightly binds to GCN4 peptide arrays (SunTag). | SunTag | Cell. 159: 635 (2014) | View |
| KRAB | Krüppel-associated box domain from human gene ZNF10 | Inhibits gene expression, commonly used with other DNA binding proteins. | Proc Natl Acad Sci U S A. 91:4509 (1994); Proc Natl Acad Sci U S A. 97: 1495 (2000) | View | |
| MS2/P65/HSF1 | Fusion protein of MS2 bacteriophage coat protein, NF-kappaB trans-activating subunit p65 and human heat-shock factor 1 activation domain | Through MS2 binding to gRNA tetraloop and stem-loop 2, p65 and HSF1 are recruited to gRNA target site and activate transcription. | EMBO J. 12:595 (1993); PLoS Biol. 7:e73 (2009); Redox Biol. 2:535 (2014) | Ver 1 Ver 2 | |
| CymR | CymR repressor | Can bind to cumate operator (CuO) sequences in the absence of cumate and blocks trascription. When cumate is added, CymR preferentially binds to it to allow transcription to proceed. | CuO | BMC Biotechnol. 6:43(2006) | View |
| CymR_NLS | CymR repressor with nuclear localization signal on both ends | Can bind to cumate operator (CuO) sequences in the absence of cumate and blocks trascription. When cumate is added, CymR preferentially binds to it to allow transcription to proceed. | CuO | BMC Biotechnol. 6:43(2006); Engineered by VectorBuilder | View |
| LacI:T2A:Neo | Lac repressor and neomycin resistance gene linked by T2A | Allows cells to express LacI protein and be resistant to neomycin. In the absence of IPTG, LacI binds to lac operator (LacO) to repress transcription of downstream genes or small RNAs. In the presence of IPTG, LacI undergoes a conformational change and is no longer able to bind to LacO, thus allowing downstream genes or small RNAs to be transcribed. | LacO | Engineered by VectorBuilder | View |
| LacI:T2A:Puro | Lac repressor and puromycin resistance gene linked by T2A | Allows cells to express LacI protein and be resistant to puromycin. In the absence of IPTG, LacI binds to lac operator (LacO) to repress transcription of downstream genes or small RNAs. In the presence of IPTG, LacI undergoes a conformational change and is no longer able to bind to LacO, thus allowing downstream genes or small RNAs to be transcribed. | LacO | Engineered by VectorBuilder | View |
| LacI:T2A:Hygro | Lac repressor and hygromycin resistance gene linked by T2A | Allows cells to express LacI protein and be resistant to hygromycin. In the absence of IPTG, LacI binds to lac operator (LacO) to repress transcription of downstream genes or small RNAs. In the presence of IPTG, LacI undergoes a conformational change and is no longer able to bind to LacO, thus allowing downstream genes or small RNAs to be transcribed. | LacO | Engineered by VectorBuilder | View |
| LacI:T2A:Bsd | Lac repressor and blasticidin resistance gene linked by T2A | Allows cells to express LacI protein and be resistant to blasticidin. In the absence of IPTG, LacI binds to lac operator (LacO) to repress transcription of downstream genes or small RNAs. In the presence of IPTG, LacI undergoes a conformational change and is no longer able to bind to LacO, thus allowing downstream genes or small RNAs to be transcribed. | LacO | Engineered by VectorBuilder | View |
人工毒蕈碱型受体
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| hM3D(Gq) | A modified form of human Gq-coupled M3 muscarinic receptor, belonging to a class of chemogenetic receptors called designer receptors exclusively activated by designer drugs (DREADD) | Allows engagement of the Gq signaling pathway in the presence of the synthetic ligand clozapine-N-oxide (CNO), leading to intracellular calcium ion release and enhanced neuronal excitation. | Proc Natl Acad Sci U S A. 104:5163 (2007) | View |
| hM4D(Gi) | A modified form of human Gi-coupled M4 muscarinic receptor, belonging to a class of chemogenetic receptors called designer receptors exclusively activated by designer drugs (DREADD) | Allows engagement of the Gi signaling pathway in the presence of the synthetic ligand clozapine-N-oxide (CNO), leading to potassium ion influx and decreased neuronal firing rates. | Proc Natl Acad Sci U S A. 104:5163 (2007) | View |
Tet调控蛋白
| 姓名 | 描述 | Application Notes | Target Sites | References | 序列 |
|---|---|---|---|---|---|
| tTA | Tetracycline transactivator | It binds to TRE promoter to activate gene transcription only in the absence of tetracycline or its analogs (e.g. doxycycline). | TRE | Proc Natl Acad Sci U S A. 89:5547 (1992) | View |
| tTA2 | Tetracycline transactivator (2nd generation) | It binds to TRE promoter to activate higher-level gene transcription in the absence of tetracycline or its analogs (e.g. doxycycline) compared to its predecessor, tTA. | TRE | Proc Natl Acad Sci U S A. 97:7963 (2000) | View |
| rtTA | Reverse tetracycline responsive transcriptional activator M2 (2nd generation) | It binds to TRE promoter to activate gene transcription only in the presence of tetracycline or its analogs (e.g. doxycycline). It has higher sensitivity to the inducing drug and lower leaky activity in the absence of the drug compared to its predecessor, rtTA. | TRE | Science. 268:1766 (1995); Proc Natl Acad Sci U S A. 97:7963 (2000) | View |
| tTS | Tetracycline transcriptional silencer | It binds to TRE promoter to actively suppress gene transcription only in the absence of tetracycline or its analogs (e.g. doxycycline). | TRE | J Gene Med. 1:4 (1999) | View |
| tTS/rtTA | tTS and rtTA_M2 linked by T2A | In the absence of tetracycline or its analogs (e.g. doxycycline), tTS binds to TRE promoter to actively suppress gene transcription whereas rtTA_M2 is unable to bind to TRE promoter. In the presence of the inducing drug, tTS dissociates from TRE promoter whereas rtTA_M2 binds to TRE promoter to activate gene transcription. Compared to using rtTA_M2 alone, which does not provide active repression, this system has lower leaky expression in the absence of the inducing drug. | TRE | Engineered by VectorBuilder based on Semin Cell Dev Biol. 13:121 (2002) | View |
| Tet3G | 3rd generation tet regulatory protein | It binds to TRE3G promoter to activate gene transcription in the presence of tetracycline or its analogs (e.g. doxycline). | TRE3G | Gene Ther. 13:1382 (2006) | View |
DNA重组酶
| 姓名 | 描述 | Application Notes | Target Sites | References | 序列 |
|---|---|---|---|---|---|
| Cre | Cyclization recombinase with SV40 nuclear localization signal | Most commonly used site-specific recombinase | LoxP and its variants (e.g. LoxN, Lox2272, Lox511, Lox66, Lox71) | Nucleic Acids Res. 32:6086 (2004); Nat Methods. 10:1028 (2013) | Ver 1 Ver 2 |
| InCre | Cre with SV40 T-antigen intron inserted in the ORF | Prevents recombinase expression in E. coli while maintaining function in mammalian cells. | LoxP and its variants (e.g. LoxN, Lox2272, Lox511, Lox66, Lox71) | Engineered by VectorBuilder | View |
| Flpo | Mouse codon-optimized version of Flpe recombinase with SV40 nuclear localization signal | Higher efficiency compared to its predecessor, Flpe, in mammalian cells. | FRT and its variants (e.g. FRT3, FRT5) | PLoS One. 2:e162 (2007); Genesis. 48:512 (2010) | View |
| InFlpo | Flpo with SV40 T-antigen intron inserted in the ORF | Prevents recombinase expression in E. coli while maintaining function in mammalian cells. | FRT and its variants (e.g. FRT3, FRT5) | Engineered by VectorBuilder | View |
| Dre | Dre Recombinase | A highly efficient site-specific recombinase similar to Cre, but recognizing different target sites. | Rox and its variants (e.g. Rox4R, Rox6R, Rox2N ) | Nucleic Acids Res. 32:6086 (2004) | View |
| InDre | Dre with SV40 T-antigen intron inserted in the ORF | Prevents recombinase expression in E.coli while maintaining function in mammalian cells. | Rox and its variants (e.g. Rox4R, Rox6R, Rox2N ) | Engineered by VectorBuilder | View |
| PBase | Native piggyBac transposase from Trichoplusia ni | Recognizes the two TRs on the piggyBac transposon and inserts the flanked region including the two TRs into host chromosomal sites that contain the TTAA sequence through a precise cut-and-paste mechanism. | PiggyBac inverted terminal repeat (ITR) | Virology. 172:156 (1989); Cell. 122:473 (2005); Nat Genet. 39:922 (2007) | View |
| hyPBase | Hyperactive version of piggyBac transposase (PBase) created by mutagenesis | Higher efficiency compared to its predecessor, PBase, in mammalian cells. | PiggyBac invertet terminal repeat (ITR) | Proc Natl Acad Sci U S A. 108:1531 (2011) | View |
| hyPBase (R372A/K375A/D450N) | Excision competent/integration defective piggyBac transposase with R372A, K375A and D450N mutations | Allows piggyBac transposon excision without potentially harmful reintegration. | PiggyBac inverted terminal repeat (ITR) | Proc Natl Acad Sci U S A. 110:E2279 (2013); J Biol Chem. 292: 6148 (2017) | View |
| Tol2 | Transposase encoded from Tol2 transposon, a member of hAT transposable element family found in medaka fish genome | Catalyzes the transposition of a non-autonomous Tol2 construct (a construct containing a deletion in the transposase coding region but retaining the Tol2 ends.) | Tol2 invertet terminal repeat (ITR) | PLoS Genet. 2:e169 (2006); Gene. 425:64 (2008); Methods Mol Biol. 561:41 (2009) | View |
| SB100X | Hyperactive variant of Sleeping Beauty transposase | Approximately 100-fold higher efficiency compared to the first-generation Sleeping Beauty transposase.Enables robust, stable gene transfer in vertebrates. | Inverted/direct repeats (IR/DR) of Sleeping Beauty transposon | Nat Genet. 41:753 (2009) | View |
| Mos1 | Drosophila Mos1 transposase inserted with an synthetic intron | Recognizes the two TRs on the Mos1 transposon and inserts the flanked region including the two TRs into host chromosomal sites that contain the TA sequence through a precise cut-and-paste mechanism. | Inverted/direct repeats (IR/DR) of Mos1 transposon | Trends Genet. 15:326 (1999); Nat Methods. 9:117 (2012) | View |
药物性DNA重组酶
| 姓名 | 描述 | Application Notes | Target Sites | References | 序列 |
|---|---|---|---|---|---|
| CreERT2 | Cyclization recombinase fused with ERT2 | Functions in the presence of tamoxifen | LoxP and its variants (e.g. LoxN, Lox2272, Lox511, Lox66, Lox71) | Proc Natl Acad Sci U S A. 104:1027 (2007) | View |
| DD-Cre | Destabilized cyclization recombinase | Functions in the presence of trimethoprim | LoxP and its variants (e.g. LoxN, Lox2272, Lox511, Lox66, Lox71) | Nat Methods. 10:1085 (2013) | View |
Cas蛋白
| 姓名 | 描述 | Application Notes | PAM Sequence | References | 序列 |
|---|---|---|---|---|---|
| zCas9 | Zebrafish codon-optimized version of S. pyogenes Cas9 with SV40 large T-antigen nuclear localization signals (nls) at both its amino and carboxyl termini | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci USA. 110: 13904 (2013) | View |
| hCas9 | Human codon-optimized CRISPR associated protein 9 from Streptococcus pyogenes (with 3xFLAG tag and nuclear localization signal), also known as SpCas9 | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Science. 339:819 (2013) | View |
| Cas9(D10A) | Human codon-optimized CRISPR associated protein 9 with D10A mutation, HA tag and nuclear localization signal | Nickase; generates 5' overhang; can induce NHEJ with paired offset gRNAs; reduced off-target effect. | NGG (preferred); NAG (less preferred) | Cell. 154: 1380 (2013) | View |
| dCas9 | Catalytically inactive Cas9, a variant of Cas9 bearing both D10A and H840A mutations | Catalytically inactive but can be recruited by gRNAs to target sites; can be fused to other proteins to direct them to specific genomic locations (e.g. dCas9 fused with transcriptional activators or repressors can lead to activation or repression of target genes). | NGG (preferred); NAG (less preferred) | Nat Biotechnol. 32: 347 (2014) | View |
| dCas9/VP64 | dCas9 fused with a VP64 acidic transactivation domain | Used for RNA-guided transcriptional activation | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 97:1495 (2000); Nat Methods. 10:973 (2013) | View |
| dCas9/VPR | dCas9 fused with a tripartite VP64-p65-Rta (VPR) transactivation domain | Used for RNA-guided transcriptional activation. | NGG (preferred); NAG (less preferred) | Nat Methods. 12:326 (2015) | View |
| dCas9/KRAB | dCas9 fused with a KRAB transcriptional silencing domain | Used for RNA-guided transcriptional repression | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 97:1495 (2000); Cell. 159:647 (2014) | View |
| dCas9/KRAB/MeCP2 | dCas9 fused with a carboxy terminal bipartite repressor domain, KRAB-MeCP2 | Efficiently suppresses gene transcription by physically blocking RNA polymerase passage by dCas9 and further transcriptional inhibition by the KRAB-MeCP2 repressor domain. | NGG (preferred); NAG (less preferred) | Cell. 88:471 (1997); Nat Methods. 15:611 (2018) | View |
| dCas9-10xGCN4_v4 | dCas9 with SV40 large T-antigen nuclear localization signals (nls) at both its amino and carboxyl termini, fused to an optimized GCN4 antibody-peptide pair (containing v4 version of the GCN4 peptide array with 10 copies of the peptide binding site, also know as SunTag10x_v4) | Modulates transcription of endogenous genes by recruitment of multiple copies of regulatory effector domains to genomic sites of interest targeted by dCas9 protein via binding of scFv antibody to a GCN4 repeating peptide array termed SunTag. | NGG (preferred); NAG (less preferred) | Cell. 159: 635 (2014) | View |
| SpCas9-HF1 | High-fidelity variant of SpCas9 | Designed to reduce non-specific DNA contacts; fewer off-target cuts. | NGG (preferred); NAG (less preferred) | Nature. 529:490 (2016) | View |
| eSpCas9 | Enhanced specificity SpCas9 variant | Low off-target effects and robust on-target cleavage. | NGG (preferred); NAG (less preferred) | Science. 351:84 (2016) | View |
| BE3 | rAPOBEC–XTEN–Cas9_D10A–UGI (rat cytidine deaminase enzyme fused with Cas9_D10A and uracil DNA glycosylase inhibitor via an XTEN linker) | Converts cytosine (C) to uracil (U) between the 4th and 8th bases of the sgRNA binding site, targeting the nonbinding strand. The modified base can be permanently converted to thymine (T) following DNA replication or repair. | NGG (preferred); NAG (less preferred) | Nature. 533:420 (2016); Nat Methods. 14:710 (2017) | View |
| Cas12a | Also known as Cpf1; single RNA-guided endonuclease of a class 2 CRISPR-Cas system from Acidaminococcus, human codon-optimized, and linked to an HA tag and nuclear localization signal | Cleaves DNA via a staggered DNA double strand break. Able to process its own CRISPR RNA (crRNA), which can simplify multiplexed genome editing. | TTTN | Cell. 163:759 (2015);Nat Biotechnol. 35:31 (2017) | View |
| Cas9/mSA | hCas9 fused with monomeric streptavidin | Used for efficient generation of targeted large insertions using biotinylated repair templates. | NGG (preferred); NAG (less preferred) | Biotechnol Bioeng. 110:57 (2013); Nat Biotechnol. 36:632 (2018) | View |
| SaCas9 | Staphylococcus aureus CRISPR associated protein 9 with HA tag and nuclear localization signal | Shorter sequence; especially useful in vector systems that have limited cargo space (e.g. AAV vectors). | NNGRR; NNGRRT(preferred) | Nature. 520:186 (2015) | View |
| hCas9:T2A:Neo | hCas9 and neomycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to neomycin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| hCas9:T2A:Puro | hCas9 and puromycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to puromycin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| hCas9:T2A:Hygro | hCas9 and hygromycin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to hygromycin B. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| hCas9:T2A:Bsd | hCas9 and blasticidin resistance gene linked by T2A | Allows cells to express hCas9 protein and be resistant to blasticidin. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| hCas9:P2A:EGFP | hCas9 and EGFP linked by P2A | Allows cells to express hCas9 protein and be visualized by green fluorescence. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| hCas9:P2A:mCherry | hCas9 and mCherry linked by P2A | Allows cells to express hCas9 protein and be visualized by red fluorescence. | NGG (preferred); NAG (less preferred) | Engineered by VectorBuilder | View |
| DmCas9 | Fruit fly (Drosophila melanogaster) codon-optimized CRISPR associated protein 9 with nuclear localization signal | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Proc Natl Acad Sci U S A. 111:E2967 (2014) | View |
| CjCas9 | A small Cas9 orthologue derived from Campylobacter jejuni with a carboxy terminal nuclear localization signal and HA tag | Shorter sequence; especially useful in vector systems that have limited cargo space (e.g. AAV vectors). | NNNNR(A/G)Y(C/T)AC; NNNNACAC (most preferred) | Nat Commun. 8:14500 (2017) | View |
| CeCas9 | S. pyogenes Cas9 codon-optimized for C. elegans, containing multiple synthetic introns, a carboxy terminal nuclear localization signal, and an HA tag | Generates double-strand DNA breaks. | NGG (preferred); NAG (less preferred) | Nat Methods. 10:1028 (2013) | View |
永生化细胞诱导基因
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| SV40-T | Simian virus 40 large T antigen | Capable of inducing mlignant transformation of a variety of cell types; can be used to immortalize primary cells. | J Virol. 64:3350 (1990) | View |
| hHRAS | Homo sapiens Ras proto-oncogene. Belongs to the Ras family of small GTPases, whose members are related to the transforming genes of mammalian sarcoma retroviruses | Mutations and overexpression of this gene can induce transformation of a variety of cell types. | Nature. 304:596 (1983); Genes Dev.15:50 (2001); Oncogene. 21:4577 (2002) | View |
| hTERT | Homo sapiens telomerase reverse transcriptase gene. Plays an improtant role in cellular senescence, and deregulation of telomerase expression in somatic cells may be involved in oncogenesis | Capable of inducing transformation of a variety of cells, and can be used to immortalize primary human cells. | Genes Dev.15:50 (2001); Cancer Res. 61:3556 (2001); Oncogene. 21:4577 (2002); PLoS Pathog.9:e1003284 (2013) | View |
| Ad5E1A | Human adenovirus type 5 E1A protein gene | Capable of inducing cellular transformation. Can be used to immortalize various primary cells. | Hum Gene Ther.11:2105 (2000); J Virol. 91:e01782 (2016) | View |
| HPV16E6 | Human papillomavirus type 16 transforming protein E6 gene | Capable of inducing cellular transformation. Can be used to immortalize various primary cells. | EMBO J.8:3905 (1989); Mol Cell J Virol. 65:4860 (1991); Biol. 24:2144 (2004) | View |
| HPV16E7 | Human papillomavirus type 16 transforming protein E7 gene | Capable of inducing cellular transformation. Can be used to immortalize various primary cells. | J. Gen. Virol. 70:1261 (1989); EMBO J.8:3905 (1989); J Virol.81:12689 (2007) | View |
| hTP53(V143A) | Homo sapiens tumor suppressor p53 gene with a V143A mutation | Can be used to induce maligant transformation of a variety of cells. | Proc Natl Acad Sci U S A. 96:8438 (1999) | View |
光敏感型蛋白
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| ChR2 | Human codon-optimized channelrhodopsin-2 from Chlamydomonas reinhardtii, a blue light-gated, cation-selective transmembrane channel protein. It rapidly undergoes conformational change upon absorbing photons to open channel permeable to cations (Na+, K+, H+, Ca2+, etc.) | Can activate excitable cells (such as neurons, muscle cells, pancreatic cells, and immune cells) by inducting depolarization of the cell membrane upon blue light stimulation; rapid ON-rate and moderate channel closing rate; maximum excitation is 470 nm. | Proc Natl Acad Sci U S A. 100:13940 (2003); Nature. 446:633 (2007); Exp Physiol.96:19 (2011) | View |
| ChR2/mCherry | ChR2 fused with mCherry by GCGGCCGCC | mCherry can mark cellular localization of ChR2; maximum excitation of ChR2 is 470 nm; maximum excitation and emission of mCherry are 587 nm and 610 nm, respectively. | Nature. 446:633 (2007) | View |
| ChR2/EYFP | ChR2 fused with EYFP by GCGGCCGCC | EYFP can mark cellular localization of ChR2; maximum excitation of ChR2 is 470 nm; maximum excitation and emission of EYFP are 513 nm and 527 nm, respectively. | Nature. 446:633 (2007) | View |
| ChR2(H134R) | A gain-of-function mutant (H134R) of human codon-optimized channelrhodopsin-2 from Chlamydomonas reinhardtii | Compared to ChR2, it has increased light sensitivity, slower channel closing rate, enhanced stationary phtocurrent and modest reduction in desensitisation, but it is less temporally precise; maximum excitation is 470 nm. | Curr Biol. 15:2279 (2005); Exp Physiol. 96:19 (2011) | View |
| ChR2(H134R)/mCherry | ChR2(H134R) fused with mCherry by GCGGCCGCC | mCherry can mark cellular localization of ChR2_H134R; maximum excitation of ChR2_H134R is 470 nm; maximum excitation and emission of mCherry are 587 nm and 610 nm, respectively. | Nature. 446:633 (2007) | View |
| ChR2(H134R)/EYFP | ChR2(H134R) fused with EYFP by GCGGCCGCC | EYFP can mark cellular localization of ChR2_H134R; maximum excitation of ChR2_H134R is 470 nm; maximum excitation and emission of EYFP are 513 nm and 527 nm, respectively. | Nature. 446:633 (2007) | View |
| eNpHR2.0/EYFP | Enhanced Halorhodopsin from Natronomonas pharaonis (NpHR) which is fused with EYFP. NpHR is a fast light-activated electrogenic Cl- pump. There is an nAChR-derived signal peptide tagged to the N-terminal of NpHR, and a Kir2.1-derived ER export signal peptide tagged to the C-terminal of EYFP to facilitate cell surface expression of NpHR | Light activation of NpHR leads to cell hyperpolarization and inhibition of firing action potentials; increased peak photocurrent compared to original NpHR; no aggregation; low cellular toxicity; maximum excitation is 580 nm; maximum excitation and emission of EYFP are 513 nm and 527 nm, respectively. | Nature. 446:633 (2007); Brain Cell Biol. 36:129 (2008) | View |
| eNpHR3.0/EYFP | Third generation of Halorhodopsin from Natronomonas pharaonis (NpHR) which is fused with EYFP. NpHR is a fast light-activated electrogenic Cl- pump. There is an nAChR-derived signal peptide tagged to the N-terminal of EYFP, and a Kir2.1-derived ER export signal peptide tagged to the C-terminal of EYFP to facilitate cell surface expression of NpHR | Light activation of NpHR leads to cell hyperpolarization and inhibition of firing action potentials; increased peak photocurrent compared to original NpHR; no aggregation; low cellular toxicity; maximum excitation is 580 nm; maximum excitation and emission of EYFP are 513 nm and 527 nm, respectively. | Front Behav Neurosci. 9: 286. (2015) | View |
| eNpHR3.0/mCherry | Third generation of Halorhodopsin from Natronomonas pharaonis (NpHR) which is fused with mCherry. NpHR is a fast light-activated electrogenic Cl- pump. There is an nAChR-derived signal peptide tagged to the N-terminal of mCherry, and a Kir2.1-derived ER export signal peptide tagged to the C-terminal of mCherry to facilitate cell surface expression of NpHR | Light activation of NpHR leads to cell hyperpolarization and inhibition of firing action potentials; increased peak photocurrent compared to original NpHR; no aggregation; low cellular toxicity; maximum excitation is 580 nm; maximum excitation and emission of mCherry are 587 nm and 610 nm, respectively. | Unpublished, provided by Karl Deisseroth through the UNC vector core | View |
钙指示剂
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| GCaMP6s | Variant of green fluorescent calcium indicator GCaMP6 with slow kinetics, ultrasensitive sensor of free calcium | Rise time: 100-150 ms, Kd = 144 nM; Most sensitive | Nature. 499:295 (2013); Cell Calcium. 58:638 (2015) | View |
| GCaMP6m | Variant of green fluorescent calcium indicator GCaMP6 with medium kinetics, ultrasensitive sensor of free calcium | Rise time: 74-100 ms, Kd = 167 nM | Nature. 499:295 (2013); Cell Calcium. 58:638 (2015) | View |
| GCaMP6f | Variant of green fluorescent calcium indicator GCaMP6 with fast kinetics, ultrasensitive sensor of free calcium | Rise time: 50-75 ms, Kd = 375 nM; Fastest | Nature. 499:295 (2013); Cell Calcium. 58:638 (2015) | View |
| jGCaMP7s | Variant of the green fluorescent protein-based calcium indicator GCaMP7 with slow kinetics; ultrasensitive sensor of free calcium | Half-rise time: 70±2 ms, Kd = 68 nM; Most sensitive | Nat Methods. 16:649 (2019) | View |
| jGCaMP7f | Variant of the green fluorescent protein-based calcium indicator GCaMP7 with fast kinetics; ultrasensitive sensor of free calcium | Half-rise time: 75±1 ms, Kd = 174 nM; Fastest | Nat Methods. 16:649 (2019) | View |
| jGCaMP7b | Variant of the green fluorescent protein-based calcium indicator GCaMP7 with brighter baseline fluorescence; ultrasensitive sensor of free calcium | Half-rise time: 80±1 ms, Kd = 82 nM; Brightest | Nat Methods. 16:649 (2019) | View |
| jGCaMP7c | Variant of the green fluorescent protein-based calcium indicator GCaMP7 which offers high contrast with low baseline fluorescence; ultrasensitive sensor of free calcium | Half-rise time: 85±3 ms, Kd = 298 nM; High contrast | Nat Methods. 16:649 (2019) | View |
| jGCaMP8s | Variant of the green fluorescent protein-based calcium indicator GCaMP8 with the highest sensitivity, suitable for in vivo imaging | Half-rise time: 10.2±0.9 ms, Kd = 46±1 nM; Fast rise, slow decay, most sensitive. | Nature. 615:884 (2023) | View |
| jGCaMP8m | Variant of the green fluorescent protein-based calcium indicator GCaMP8 with medium sensitivity and kinetics, suitable for in vivo imaging | Half-rise time: 7.4±0.6 ms, Kd = 108±3 nM; Fast rise, medium decay, moderate sensitivity. | Nature. 615:884 (2023) | View |
| jGCaMP8f | Variant of the green fluorescent protein-based calcium indicator GCaMP8 with the fastest kinetics and lower sensitivity, suitable for in vivo imaging | Half-rise time: 6.6±1 ms, Kd = 334±18 nM; Fast rise, fast decay. | Nature. 615:884 (2023) | View |
酵母筛选标记
| 姓名 | 描述 | Auxotrophic Complementation | Application Notes | References | 序列 |
|---|---|---|---|---|---|
| URA3 | Orotidine-5'-phosphate decarboxylase from the yeast Saccharomyces cerevisiae | Uracil and uridine | Allows yeast lacking endogenous URA3 gene to survive without uracil and uridine. | Mol Gen Genet. 197:345 (1984); Genetics. 122:19 (1989) | View |
| TRP1 | Phosphoribosylanthranilate isomerase TRP1 from the yeast Saccharomyces cerevisiae | Tryptophan | Allows yeast lacking endogenous TRP1 gene to survive without tryptophan. | Genetics. 122:19 (1989) | View |
| HIS3 | Imidazoleglycerol-phosphate dehydratase from the yeast Saccharomyces cerevisiae | Histidine | Allows yeast lacking endogenous HIS3 gene to survive without histidine. | Genetics. 122:19 (1989) | View |
| LEU2 | 3-isopropylmalate dehydrogenase from the yeast Saccharomyces cerevisiae | Leucine | Allows yeast lacking endogenous LEU2 gene to survive without leucine. | Genetics. 122:19 (1989) | View |
iPS干细胞诱导基因
| 姓名 | 描述 | Application Notes | References | 序列 |
|---|---|---|---|---|
| hMYC | Homo sapiens c-myc proto-oncogene. Belongs to myelocytomatosis (Myc) family of transcription factors. Plays a role in cell cycle procession, apotosis and cellular transformation | Mutations, overexpression, rearrangement and translocation of this gene can induce transformation or immortalization of a variety of cell types. | Nature. 306:194 (1983); Proc Natl Acad Sci U S A. 97:11198 (2000); Cancer Res. 65:2179 (2005); Mol Cancer Res. 5:1181 (2007); J Virol.81:12689 (2007) | View |
| hOct4 | Homo sapiens Oct-4 gene. Encodes a transcription factor containing a POU homeodomain that plays a key role in embryonic development and stem cell pluripotency. Aberrant expression in adult tissues is associated with tumorigenesis. | Required for the maintenance of stem cell properties and malignant progression in various cancers. Can be used to reprogram a variety of cell types. | Cell Research. 12:321 (2002); Science. 318:1917 (2007); J Stem Cells Regen Med. 6:149 (2010); DNA Cell Biol. 36:1000 (2017) | View |
| hSOX2 | Homo sapiens transcription factor SOX-2 gene. Belongs to the SRY-related HMG-box (SOX) family of transcription factors, which is involved in the regulation of embryonic development and in the determination of cell fate | Required for the maintenance of stem cell properties and malignant progression in various cancers. Can be used to reprogram a variety of cell types. | Science. 318:1917 (2007); J Stem Cells Regen Med. 6:149 (2010); DNA Cell Biol. 36:1000 (2017) | View |
| mMyc | Mus musculus myc proto-oncogene. Belongs to myelocytomatosis (Myc) family of transcription factors. Plays a role in cell cycle procession, apotosis and cellular transformation | Mutations, overexpression, rearrangement and translocation of this gene can induce transformation or immortalization of a variety of cell types. | Cell. 39:339 (1984); Nature. 320:760 (1986); Mol Cell Biol. 7:3899 (1987); Cell. 126:663 (2006); Nature. 451:141 (2008) | View |
| mPou5f1 | Mus musculus Oct-4 gene. Encodes a transcription factor containing a POU homeodomain that plays a key role in embryonic development and stem cell pluripotency. | Required for the maintenance of stem cell properties. Can be used to reprogram a variety of cell types. | Cell. 126:663 (2006); Nature. 448:318 (2007); Nature. 451:141 (2008) | View |
| mSox2 | Mus musculus transcription factor SOX-2 gene. Belongs to the SRY-related HMG-box (SOX) family of transcription factors, which is involved in the regulation of embryonic development and in the determination of cell fate | Required for the maintenance of stem cell properties. Can be used to reprogram a variety of cell types. | Cell. 126:663 (2006); Nature. 448:318 (2007); Nature. 451:141 (2008) | View |
| hNanog | Homo sapiens nanog homeobox gene. Encodes a DNA-binding homeobox-family transcription factor involved in embryonic stem (ES) cell proliferation, renewal, and pluripotency. | Required for the maintenance of stem cell properties and malignant progression in various cancers. Can block ES cell differentiation, autorepress its own expression in differentiating cells, and can be used to reprogram a variety of cell types. | Science. 318:1917 (2007); J Stem Cells Regen Med. 6:149 (2010); Mol Cell Biochem. 351:109 (2011); DNA Cell Biol. 36:1000 (2017) | View |
| mNanog | Mus musculus nanog homeobox gene. Encodes a DNA-binding homeobox-family transcription factor involved in embryonic stem (ES) cell proliferation, renewal, and pluripotency. | Required for the maintenance of stem cell properties. Can block ES cell differentiation, autorepress its own expression in differentiating cells, and can be used to reprogram a variety of cell types. | Cell. 113:631 (2003); Nature. 448:318 (2007) | View |
| hKLF4 | Homo sapiens Kruppel-like factor 4 (Klf4) gene. Belongs to the relatively large family of SP1-like transcription factors and is involved in the regulation of proliferation, differentiation, apoptosis and somatic cell reprogramming. | Required for the maintenance of stem cell properties. It can prevents ES cell differentiation and maintains ES cell self-renewal and pluripotency. Can be used to reprogram a variety of cell types. Can function as either an oncogene or a tumor suppressor depending on differing cellular contexts and cancer types. | J Biol Chem. 285:9180 (2010); Nat Commun. 9:1261 (2018) | View |
| mKlf4 | Mouse Kruppel-like factor (Klf4) gene. Belongs to the relatively large family of SP1-like transcription factors and is involved in the regulation of proliferation, differentiation, apoptosis and somatic cell reprogramming. | Required for the maintenance of stem cell properties. It can prevents ES cell differentiation and maintains ES cell self-renewal and pluripotency. Can be used to reprogram a variety of cell types. Can function as either an oncogene or a tumor suppressor depending on differing cellular contexts and cancer types. | J Biol Chem. 285:9180 (2010); Gene. 611: 27 (2017); Nat Commun. 9:1261 (2018) | View |
| hLIN28A | Human lin-28 homolog A gene. Encodes a LIN-28 family RNA-binding protein that acts as a posttranscriptional regulator of genes involved in development, self-renewal of embryonic stem cells and metabolism. Overexpressed in human embryonic stem cells, primary human tumors and human cancer cell lines. | It is dispensable for maintaining pluripotency. Regulate the self-renewal of stem cells. Can be used to promote efficient reprogramming of a variety of cell types. Enhances tissue repair in some adult tissues by reprogramming cellular bioenergetics. Promotes the expression of some metabolic enzymes. Can regulate glucose homeostasis in mammals and promote resistance to high fat diet-induced obesity and type 2 diabetes. | Cell. 155: 778 (2013); Cell Stem Cell. 19:66 (2016) | View |
| mLin28 | Mouse lin-28 homolog A gene. Encodes a LIN-28 family RNA-binding protein that acts as a posttranscriptional regulator of genes involved in development, self-renewal of embryonic stem cells and metabolism. Highly expressed in mouse embryonic stem cells and during early embryogenesis. | It is dispensable for maintaining pluripotency. Can be used to promote efficient reprogramming of a variety of cell types. Regulate the self-renewal of stem cells. Overexpression in mice can cause gigantism and a delay in puberty onset. Enhances tissue repair in some adult tissues by reprogramming cellular bioenergetics. Can promotes the expression of some metabolic enzymes. Can regulate glucose homeostasis in mammals and promote resistance to high fat diet-induced obesity and type 2 diabetes. | Cell. 155: 778 (2013); Cell Stem Cell. 19:66 (2016) | View |
| hESRRB | Human estrogen-related-receptor β gene. Encodes a member of the nuclear orphan receptor family that is involved in early development, pluripotency and reprogramming. | Activates Oct4 transcription and sustains self-renewal and pluripotency in embryonic stem cells. Can be used to reprogram a variety of cell types. | J Biol Chem. 283:35825 (2008); FEBS Lett. 592:852 (2018); J Cell Physiol. 233:1601 (2018) | View |
| mEsrrb | Mouse estrogen-related-receptor β gene. Encodes a member of the nuclear orphan receptor family that is involved in early development, pluripotency and reprogramming. | Activates Oct4 transcription and sustains self-renewal and pluripotency in embryonic stem cells. Can be used to reprogram a variety of cell types. | J Biol Chem. 283:35825 (2008); FEBS Lett. 592:852 (2018); J Cell Physiol. 233:1601 (2018) | View |
| mOSKM | A combination of four reprogramming factor genes (mouse Oct3/4, Sox2, Klf4 and c‐Myc) linked by three 2A “self-cleaving” peptides. Involved in reprogramming, or generation of induced pluripotent stem (iPS) cells. Their expression commonly increases in metastatic cancers. | Required for the maintenance of stem cell properties. It can prevents ES cell differentiation and maintains ES cell self-renewal and pluripotency. Can be used to reprogram both embryonic and adult somatic cells. | Stem Cells Int. 2016:9451492 (2016); Biochim Biophys Acta Mol Cell Res. 1864:1359 (2017) | View |
| hOSKM | A combination of four reprogramming factor genes (human Oct4, Sox2, Klf4 and c‐Myc) linked by three 2A “self-cleaving” peptides. Involved in reprogramming, or generation of induced pluripotent stem (iPS) cells. Their expression commonly increases in metastatic cancers. | Required for the maintenance of stem cell properties. It can prevents ES cell differentiation and maintains ES cell self-renewal and pluripotency. Can be used to reprogram both embryonic and adult somatic cells. | Stem Cells Int. 2016:9451492 (2016); Stem Cells Int. 2019: 1393791 (2019) | View |