Article Dans Une Revue STAR Protocols Année : 2023

Optimized protocol to generate genome-wide inactivated Cas9-expressing murine T cells

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In vivo genome-wide CRISPR screens in primary T cells allow the systematic and unbiased identification of non-redundant regulatory mechanisms shaping immune responses. Here, we present an optimized protocol for efficient generation of a pool of genome-wide inactivated Cas9-expressing T cells using a retroviral library of sgRNA. We detail the process of large-scale viral production and library integration in activated murine T cells as well as the two-step PCR approach for sgRNA recovery and abundance evaluation.

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hal-04968736 , version 1 (26-02-2025)

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Marguerite Laprie-Sentenac, Clara Cretet-Rodeschini, Laurie Menger. Optimized protocol to generate genome-wide inactivated Cas9-expressing murine T cells. STAR Protocols, 2023, 4, ⟨10.1016/j.xpro.2022.101922⟩. ⟨hal-04968736⟩
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