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BRIEF RESEARCH REPORT article

Front. Immunol.
Sec. T Cell Biology
Volume 15 - 2024 | doi: 10.3389/fimmu.2024.1445341

Efficient gene deletion of Integrin alpha 4 in primary mouse CD4 T cells using CRISPR RNA pair-mediated fragmentation

Provisionally accepted
Taeuk Wi Taeuk Wi 1,2Yurim Choi Yurim Choi 1,2Jungsun Kim Jungsun Kim 1,2Youn Soo Choi Youn Soo Choi 3,4Matthew E. Pipkin Matthew E. Pipkin 5Jinyong Choi Jinyong Choi 1,2*
  • 1 Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea
  • 2 Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea
  • 3 Department of Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea
  • 4 Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Republic of Korea
  • 5 Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and Technology, Jupiter, Florida, United States

The final, formatted version of the article will be published soon.

    The functional specialization of CD4 T lymphocytes into various subtypes, including TH1 and TFH cells, is crucial for effective immune responses. TFH cells facilitate B cell differentiation within germinal centers, while TH1 cells are vital for cell-mediated immunity against intracellular pathogens. Integrin α4, a cell surface adhesion molecule, plays significant roles in cell migration and co-stimulatory signaling. In this study, we investigated the role of Integrin α4 in regulating TFH and TH1 cell populations during acute viral infection using CRISPR-Cas9 gene editing. To effectively delete the Itga4 in primary mouse CD4 T cells, we selected various combinations of crRNAs and generated ribonucleoprotein complexes with fluorochrome-conjugated tracrRNAs and Cas9 proteins. These crRNA pairs enhanced gene deletion by generating deletions in the gene. By analyzing the effects of Itga4 deficiency on TFH and TH1 cell differentiation during acute LCMV infection, we found that optimized crRNA pairs significantly increased the TH1 cell population. Our results highlight the importance of selecting and combining appropriate crRNAs for effective CRISPR-Cas9 gene editing in primary CD4 T cells. Additionally, our study demonstrates the role of Integrin α4 in regulating the differentiation of CD4 T cells, suggesting the potential molecular mechanisms driving T cell subset differentiation through integrin targeting.

    Keywords: Integrin α4, CRISPR-Cas9 gene editing, TFH, Th1, viral infection

    Received: 07 Jun 2024; Accepted: 18 Nov 2024.

    Copyright: © 2024 Wi, Choi, Kim, Choi, Pipkin and Choi. 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: Jinyong Choi, Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.