Improving CRISPR Performance Through Guide RNA Enhancements
Originally aired on June 13, 2018
Technology for incorporation of chemical modifications into CRISPR synthetic guide RNAs and extending the lifetime of these reagents in transfected cells, which in turn increases editing yields, is of growing importance in clinical research, for example in genome editing applications of the sickle-cell mutation in human adult stem cells.
In this webinar, we will describe the production of an optimal, chemically protected guide for precise targeting of the sickle mutation using Cas9 ribonucleoprotein electroporation. We identified two locations in the gRNA recognition sequence for Cas9 where a 3′ PACE modification significantly enhances the on-target cleavage specificity for various clinically relevant genes. We will show how to tailor the specificity of gRNAs for select targets and cellular environments to enhance the performance of CRISPR-based editing in transfected cells.