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Our AAV CRISPR vector is designed to work with SaCas9 derived from
Staphylococcus aureus, which is >1 kb shorter in comparison to the
conventional SpCas9 derived from Streptococcus pyogenes. SaCas9 provides a
distinct advantage over SpCas9 which has limited use in AAV-based applications
due to its large size and the small cargo capacity of AAV vectors....
An AAV vector is first constructed as a plasmid in E. coli where a LoxP-
flanked gene of interest is cloned in-between two inverted terminal repeats
(ITRs). It is then transfected into packaging cells along with helper
plasmids, where the region of the vector between the two inverted terminal
repeats (ITRs) is packaged into live virus. When AAV virus is added to target
cells, gene expression can be inactivated in the presence of Cre recombinase
upon Cre-mediated excision of the coding sequence....
It is then transfected into packaging cells along with helper
plasmids, where the region of the vector between the two inverted terminal
repeats (ITRs) is packaged into live virus. The non-coding RNA of interest
placed in-between the two ITRs is introduced into target cells along with the
rest of viral genome.
The wild-type AAV genome is a linear single-stranded DNA (ssDNA) with two ITRs
forming a hairpin structure on each end. It is therefore also known as ssAAV....
It is then transfected into packaging cells along with helper
plasmids, where the region of the vector between the two inverted terminal
repeats (ITRs) is packaged into live virus. The tetracycline inducible gene
expression cassette consisting of the TRE promoter driving the user-selected
GOI is placed in-between the two ITRs, which is introduced into target cells
along with the rest of viral genome....
It is then
transfected into packaging cells along with helper plasmids, where the region
of the vector between the two inverted terminal repeats (ITRs) is packaged
into live virus. For the AAV (FLEX) conditional Cre-Switch gene expression
vector, the FLEX Cre-Switch described above is placed in-between the two ITRs
during vector construction, which is introduced into target cells along with
the rest of viral genome....
It is then transfected into
packaging cells along with helper plasmids, where the Tet regulatory protein
expression cassette between the two inverted terminal repeats (ITRs) is
packaged into active virus. Any gene(s) placed in-between the two ITRs are
introduced into target cells along with the rest of viral genome....
It is then transfected into packaging cells along with helper plasmids, where
the region of the vector between the two inverted terminal repeats (ITRs) is
packaged into live virus. The Cas9 expression cassette placed in-between the
two ITRs is introduced into target cells along with the rest of viral genome.
A user-selected promoter drives Cas9 expression, which is then directed to the
DNA target site of interest in the presence of the target site-specific gRNA
sequence....
It is then transfected into packaging cells along with helper plasmids,
where the region of the vector between the two inverted terminal repeats
(ITRs) is packaged into live virus. The donor sequence and homology arms
placed in-between the two ITRs are introduced into target cells along with the
rest of viral genome. Cas9 and gRNA from at least one other vector facilitate
a targeted DSB, allowing the donor sequence to be inserted into the host
genome....
## 概述
Recombinant adenoviral vectors are used for achieving high levels of transgene
expression in a wide variety of mammalian cell types, where the vector remains
as episomal DNA without integrating into the host genome. High transduction
efficiency and high levels of short-term gene expression make adenoviral
vectors the preferred gene delivery tools for gene therapy and vaccine
research.
Adenoviral vectors are derived from adenovirus, which causes the common cold....
It is then
transfected into packaging cells along with helper plasmids, where the region
of the vector between the two inverted terminal repeats (ITRs) is packaged
into live virus. Any gene(s) placed in-between the two ITRs are introduced
into target cells along with the rest of viral genome.
The wild-type AAV genome is a linear single-stranded DNA (ssDNA) with two ITRs
forming a hairpin structure on each end. It is therefore also known as ssAAV....
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