Rapid and efficient generation of antigen-specific isogenic T cells from cryopreserved blood samples.

ABSTRACT: Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-mediated gene editing has been leveraged for the modification of human and mouse T cells. However, limited experience is available on the application of CRISPR/Cas9 electroporation in cryopreserved T cells collected during clinical trials. To address this, we aimed to optimize a CRISPR/Cas9-mediated gene editing protocol compatible with peripheral blood mononuclear cells (PBMCs) samples routinely produced during clinical trials. PBMCs from healthy donors were used to generate knockout T-cell models for interferon-γ, Cbl proto-oncogene B (CBLB), Fas cell surface death receptor (Fas) and T-cell receptor (TCRαβ) genes. The effect of CRISPR/Cas9-mediated gene editing on T cells was evaluated usi

SUBMITTER: Eerkens AL 

PROVIDER: S-EPMC9314923 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature