PERSPECTIVE article
Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1589729
This article is part of the Research TopicSafety and efficacy of CRISPR/Cas-based genome editing tools: applications and considerations in cell and gene therapyView all articles
Strategies for CRISPR-based knock-ins in primary human B cells and lymphoma cell lines
Provisionally accepted
- 1University Hospital Frankfurt, Frankfurt, Germany
- 2MRC Cambridge Stem Cell Institute, Cambridge, United Kingdom
- 3National Institutes of Health (NIH), Bethesda, Maryland, United States
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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-Cas9based 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 cancerrelated 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.
Keywords: gene editing (CRISPR/Cas9), knock-in, Lymphoma, DLBCL, NF-kappa B (NF-KB)
Received: 07 Mar 2025; Accepted: 18 Apr 2025.
Copyright: © 2025 Lund, Gong, Yu, Staudt, Hodson and Scheich. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Sebastian Scheich, University Hospital Frankfurt, Frankfurt, Germany
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