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Updated: May 18, 2026

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines
Published on: November 13, 2021
Re-balancing immunity with CRISPR-Cas9: Novel strategies for cancer and autoimmune disorders
Xuanqi Ren1, Ziyang Zhou1, Lingkai Kong2
1Jinan Microecological Biomedicine Shandong Laboratory, Building 1, Jinan Medical and Health Science and Technology Innovation Industrial Park, No. 288, Jiqi Road, Huayin District, Jinan City, Shandong Province, China; State Key Laboratory of Pharmaceutical Biotechnology, Suqian Scientific Research Institute of Nanjing University Medical School, Division of Hepatobiliary and Transplantation Surgery, Department of General Surgery Nanjing Drum Tower Hospital, Nanjing Drum Tower Hospital, the Affiliated Hospital of Medical School, Medical School, Nanjing University, Nanjing 210008, China; "Nanjing University-Gulou" Joint Laboratory of AI and Healthcare BigData, National Institute of Healthcare Data Science at Nanjing University, School of Life Sciences, Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing 210093, China.
Abstract:
Dysregulated immune activity is a cornerstone of numerous diseases, where excessive suppression enables cancer progression and uncontrolled activation drives autoimmunity. Immunotherapy aims to restore this balance, however, conventional approaches are often limited by suboptimal precision, efficacy, or long-term safety. The emergence of the CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9) genome-editing system has opened transformative avenues for overcoming these hurdles. This review traces the development of the CRISPR-Cas9 toolkit, encompassing gene knock out (CRISPR-KO), knock in (CRISPR-KI), transcriptional activation (CRISPRa), interference (CRISPRi), base editing and prime editing-and examines their application in reprogramming immune responses. We focus on how these six strategies enhance anti-tumor immunity by engineering immune cells to bypass inhibitory checkpoints and the tumor microenvironment, and suppress autoimmunity by restoring immune tolerance through precise genomic and transcriptional interventions. Finally, we discuss translational challenges and future directions, including the adoption of next-generation editors and smart delivery systems, which are poised to maximize the therapeutic potential of CRISPR-based cellular interventions.
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