Optimized protocol for gene editing in adipocytes using CRISPR-Cas9 technology

Abstract

Summary: We present a detailed protocol for gene editing in adipocytes using the CRISPR-Cas technology. This protocol describes sgRNA design, preparation of lentiCRISPR-sgRNA vectors, functional validation of sgRNAs, preparation of lentiviruses, and lentiviruses transduction in adipocytes. Moreover, an optimized method of gene editing using the lentiCRISPRv2 vector expressing two sgRNAs targeting two different genes has also been described.For complete details on the use and execution of this protocol, please refer to Qiu et al. (2020).

Keywords

• Cell culture
• CRISPR
• Metabolism
• Molecular biology