...high efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...positive cells Packaging and envelope plasmids provide the necessary components to make lentiviral particles...amplified, PCR products are denatured-renatured, treated with a nuclease that cleaves DNA heteroduplexes...particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only compatible with smaller Cas enzymes... between your gRNA targeting sequence and target DNA may result in reduced cleavage. The number of alleles...increase the likelihood that a nonfunctional protein product is produced. Alternatively, gRNAs can be designed... is absolutely necessary for Cas9 to bind target DNA. As such, one can start by identifying all PAM sequences...

...is its single-stranded DNA genome. Because the virus depends on the cell’s DNA replication machinery to...for host cell DNA synthesis. Unfortunately, this technique further limits the packaging capacity of AAV...2.4 kb. Methods to Increase Packaging Capacity: To increase the packaging capacity of AAV, a longer transgene...pair to allow for synthesis of the complementary DNA strand. Rep and Cap are translated to produce multiple...and other optogenetic genes enables them to be packaged in AAVs. Click here for more information on optogenetics...indicates a virus containing the genome of serotype 2 packaged in the capsid from serotype 5. Use of these pseudotyped...mentioned above, AAV improvements have included the production of synthetic capsids and the mixing of capsids...

...purify specific regions of DNA, image DNA in live cells, and precisely edit DNA and RNA. The ease of generating... two formats: as liquid DNA, or in select cases, as pre-made lentivirus . DNA libraries are shipped at...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence...will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...strands of the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...of target DNA. Cas9n increases target specificity. Since two nickases targeting opposite DNA strands are...groove and the non-target DNA strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9...