Abstract
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. For complete details on the use and execution of this protocol, please refer to Sutra Del Galy et al. (2020).
Original language | English |
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Article number | 101922 |
Journal | STAR Protocols |
Volume | 4 |
Issue number | 1 |
DOIs | |
Publication status | Published - 17 Mar 2023 |
Keywords
- CRISPR
- Immunology