Manufacturing of CRISPR-edited primary mouse CAR T cells for cancer immunotherapy



Summary

CRISPR-Cas9 technology can enhance CAR T-cell immunotherapy by improving T-cell function and targeting specificity. This study focuses on a streamlined manufacturing process for generating CRISPR-edited, primary mouse CAR T cells. The process involves electroporation of Cas9 ribonucleoprotein (RNP) complexes into freshly isolated T cells to knock out target genes, followed by lentiviral transduction to introduce the CAR construct. Crucially, the process avoids prolonged in vitro activation, preserving T-cell functionality. This method demonstrates efficient gene editing and CAR expression, resulting in CAR T cells that exhibit superior anti-tumor activity in vivo compared to traditional CAR T cells, paving the way for improved cancer therapies.

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