...System Design gRNA Know your Cell Line Select Gene Select gRNA Clone gRNA Delivery of gRNA & Cas9 Validate...a guide RNA (gRNA or sgRNA) and a CRISPR-associated endonuclease (Cas protein). The gRNA is a short synthetic...how to design your gRNA . Browse Plasmids: gRNAs Synthesize and clone desired gRNAs Once you have selected...commonly referred to as an sgRNA. gRNA scaffold sequence The sequence within the gRNA that is responsible for...protein and the gRNA form a ribonucleoprotein complex through interactions between the gRNA scaffold and ...the gRNA remains free to interact with target DNA. Cas9 will only cleave a given locus if the gRNA spacer...designing a gRNA for your experiment (see Plan Your Experiment below). In addition to optimizing gRNA design...

...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...

...planning your CRISPR experiment and designing your gRNA and how to use pooled libraries in genome-wide screens...

...γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA of interest: The transgene sequence is...

...also transcribes small RNAs, such as shRNAs and gRNAs. Resources Addgene's blog , including our popular...