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

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

STAR protocols (2021-02-09)
Yan Qiu, Qiurong Ding
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Calcium chloride dihydrate, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Millipore
RIPA Lysis Buffer, 10X, 100 mL RIPA Lysis Buffer, 10X for Immunoprecipitation & Western Blotting.