Optimized methods for using Cas9 nickases in genome editing
The CRISPR-Cas9 system has emerged as one of the leading tools for modifying genomes of organisms ranging from E. coli to humans. One of the key components of this editing system is Cas9 endonuclease, and a double nicking strategy can be leveraged to reduce unwanted off-target effects. However, the nickase experiments can be inherently more complicated than standard CRISPR-Cas9 editing, given the requirement for two guide RNAs to function simultaneously. In this webinar, both Shuqi Yan and Mollie Schubert present data from the characterization of various factors that impact the efficiency of cooperative nicking in cell cultures. They also summarize key design considerations for efficient gene disruption or homology directed repair (HDR) when planning your nickase experiments.