Efficient Generation of Gene-edited Mouse Models and Cell Lines Using Synthetic sgRNA

The CRISPR/Cas genome editing system has revolutionized nearly every aspect of the life science industry. Until recently, the most used formats for this technology have been plasmids, mRNA, or lentivirus. Synthetic single modified guide RNA (sgRNA) delivered pre-complexed with Cas nucleases reduces the limiting factors of the above formats such as off-target effects, low efficiency, and undesired cellular responses to foreign DNA.  Discover how the Genome Editing Shared Resource at Rutgers Cancer Institute vetted and validated the transition from in vitro transcribed CRISPR sgRNA/Cas9 mRNA to synthetic two-part CRISPR RNA:trans-activating CRISPR RNA (crRNA:tracrRNA) and Cas9 protein and then to full-length synthetic sgRNA ribonucleoprotein (RNP) complexes. Applications of sgRNA in animal model generation and cell line applications will be discussed.

Learning Objectives:

1. Best practices for using synthetic sgRNA for mouse model generation

2. How to use sgRNA RNPs for a variety of cell-based applications