- 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
PMID33554142
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).
MATERIALI
N° Catalogo
Marchio
Descrizione del prodotto
Millipore
Tampone di lisi RIPA, 10X, 100 mL RIPA Lysis Buffer, 10X for Immunoprecipitation & Western Blotting.
Sigma-Aldrich
Cloruro di calcio, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%