We narrowed to 9 results for: grn
-
TypeGuide... guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme). The gRNA is a short synthetic...conversions using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target site... of unique gRNAs. gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high ...commonly referred to as an sgRNA. gRNA scaffold sequence The sequence within the gRNA that is responsible for...amplify a specific sequence of DNA pegRNA P rime E dit gRNA; a specialized gRNA containing an additional primer...Cas enzyme and a gRNA specific to the gene of interest. The genomic target of the gRNA can be any ~20 nucleotide...interactions between the gRNA scaffold and surface-exposed positively-charged grooves on Cas9. gRNA binding induces...
-
Plan Your Experiment
TypeGuide...edit type, designing and cloning a gRNA, delivering Cas and the gRNA, and validating the edit.... system for Cas9 and the guide RNA (gRNA), the design of the gRNA sequence, and finally how to validate... proper gRNA design. While there are some cases where it makes sense to manually design a gRNA, in most...most cases gRNA design software is better and more efficient at designing potential gRNAs for you to choose...testing multiple gRNAs is a good idea for most CRISPR experiments, testing multiple pegRNA designs is particularly...your gRNA is the balance between on-target and off-target activity. In a perfect world, your gRNA sequence...mismatches between a given gRNA sequence and your genomic target may reduce the gRNA activity. Be sure to sequence... -
Guide to Using Pooled Libraries
TypeGuide...each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains a unique gene targeting...library from the Moffat lab has over 175,000 different gRNA-containing plasmids). Amplifying and Using the Library... screen. The figure above is an outline of how a gRNA library is used. Below are the general steps to ...experimental and control cells Generate a list of gRNAs that disappear with addition of the selection mechanism... -
Lentiviral Vector Guide
TypeGuide... require gRNAs for each target. Lentiviral vectors are a common way of delivering these gRNAs, due to ...plasmids: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; 8–10 kb packaging...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ... -
Gamma-Retroviral Vector Guide
TypeGuide...Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; ~8 kb packaging...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ... -
Adenovirus Guide
TypeGuide...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ...capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral particle... -
Adeno-associated virus (AAV) Guide
TypeGuide...Vector Production Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid ... Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV capsid is challenging due ... -
Addgene's eBook Collection
TypeGuide...planning your CRISPR experiment and designing your gRNA and how to use pooled libraries in genome-wide screens... -
Promoters
TypeGuide...also transcribes small RNAs, such as shRNAs and gRNAs. Resources Addgene's blog , including our popular...