Microinjection for precision genome editing in Caenorhabditis elegans

Abstract

Summary: In Caenorhabditis elegans, targeted genome editing techniques are now routinely used to generate germline edits. The remarkable ease of C. elegans germline editing is attributed to the syncytial nature of the pachytene ovary which is easily accessed by microinjection. This protocol describes the step-by-step details and troubleshooting tips for the entire CRISPR-Cas genome editing procedure, including gRNA design and microinjection of ribonucleoprotein complexes, followed by screening and genotyping in C. elegans, to help accessing this powerful genetic animal system.For complete details on the use and execution of this protocol, please refer to Ghanta and Mello (2020).

Keywords

• genetics
• model organisms
• molecular biology
• CRISPR