TY  - JOUR
AU  - Lund, Sophie
AU  - Gong, Chun
AU  - Yu, Xin
AU  - Staudt, Louis M
AU  - Hodson, Daniel J
AU  - Scheich, Sebastian
TI  - Strategies for CRISPR-based knock-ins in primary human B cells and lymphoma cell lines.
JO  - Frontiers in immunology
VL  - 16
SN  - 1664-3224
CY  - Lausanne
PB  - Frontiers Media
M1  - DKFZ-2025-01131
SP  - 1589729
PY  - 2025
AB  - Since its advent about ten years ago, the CRISPR-Cas9 system has been frequently used in biomedical applications. It has advanced various fields, and CRISPR-Cas9-based therapeutics have shown promising results in the treatment of specific hematological diseases. Furthermore, CRISPR gene editing technologies have revolutionized cancer research by enabling a broad range of genetic perturbations, including genetic knockouts and precise single nucleotide changes. This perspective focuses on the state-of-the-art methodology of CRISPR knock-ins to engineer immune cells. Since this technique relies on homology-directed repair (HDR) of double-strand breaks (DSBs) induced by the Cas9 enzyme, it can be used to introduce specific mutations into the target genome. Therefore, this methodology offers a valuable opportunity to functionally study specific mutations and to uncover their impacts not only on overall cell functions but also on the mechanisms behind cancer-related alterations in common signaling pathways. This article highlights CRISPR knock-in strategies, protocols, and applications in cancer and immune research, with a focus on diffuse large B cell lymphoma.
KW  - Humans
KW  - CRISPR-Cas Systems
KW  - Gene Knock-In Techniques: methods
KW  - Gene Editing: methods
KW  - B-Lymphocytes: metabolism
KW  - B-Lymphocytes: immunology
KW  - Cell Line, Tumor
KW  - Lymphoma, Large B-Cell, Diffuse: genetics
KW  - DLBCL (Other)
KW  - NF-kappa B (NF-KB) (Other)
KW  - gene editing (CRISPR/Cas9) (Other)
KW  - knock-in (Other)
KW  - lymphoma (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:40453079
C2  - pmc:PMC12122508
DO  - DOI:10.3389/fimmu.2025.1589729
UR  - https://inrepo02.dkfz.de/record/301747
ER  -