CRISPR-Cas9 is a potent and easily reprogrammable tool for RNA-targeted induction of gene knockouts. Designing guide RNA (gRNA) to create double-strand breaks (DSB) at a specific genomic location can seem straightforward (e.g. simply look for a PAM site),but there are many factors to consider for ensuring that the genome editing occurs precisely at the desired locus while avoiding off-target effects.

In this webinar, we will describe the development of the Dharmacon™ Edit-R™ CRISPR algorithm and the selection of predesigned guide RNAs for functional and highly specific knockouts. Using more than 25 years of experience chemically synthesizing RNA and creating innovative Dharmacon Reagents applications, we have synthesized more than 1,200 CRISPR-Cas9 gRNAs and tested them in a reporter cell line assay for functional gene knockout. We next combined this direct knowledge of potent sequences with a groundbreaking, exhaustive mismatch search tool into an algorithm to coordinately select both the most active and most sequence-specific gRNAs. These gRNAs are guaranteed to affect gene disruption and ensure functional gene product knockout. We validated the predesigned knockout guides in ELISA, immunofluorescence, and other assays, including in primary human immune cells. Potent and specific Edit-R CRISPR knockout guides are an effective tool for disease modeling in cell lines, for screening of gene function, and as a complement to other RNA-guided knockdown technologies such as CRISPR interference (CRISPRi) and small interfering RNA (siRNA).

In this webinar, you will gain insights about:

·  The importance of using highly functional and specific CRISPRko gRNAs

·  Rational development of the Dharmacon Edit-R CRISPRko algorithm

·  Extensive algorithm validation to ensure universal phenotypic outcomes of predicted guides, in many cell lines including primary cells