Production of CRISPRi-engineered primary human mammary epithelial cells with baboon envelope pseudotyped lentiviral vectors.
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ABSTRACT:
SUBMITTER: Pastor S
PROVIDER: S-EPMC9871349 | biostudies-literature | 2023 Mar
REPOSITORIES: biostudies-literature
Publications
Production of CRISPRi-engineered primary human mammary epithelial cells with baboon envelope pseudotyped lentiviral vectors.
Pastor Sonia S Wicinski Julien J Charafe-Jauffret Emmanuelle E Verhoeyen Els E Guittard Geoffrey G Ginestier Christophe C
STAR protocols 20230119 1
Primary human mammary epithelial cells (pHMECs) are known to be remarkably difficult to engineer genetically. Here, we present a protocol for efficient transduction of pHMECs using a baboon retroviral envelope glycoprotein for pseudotyping of lentiviral vectors (BaEV-LVs). We describe the preparation of the BaEV-LVs, the isolation of pHMECs from breast samples, and the subsequent transduction of pHMECs. We also detail the use of CRISPRi technology to efficiently silence gene expression in pHMECs ...[more]
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