ORIGINAL RESEARCH article
Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
FACS-Based Genome-Wide CRISPR Screening Platform Identifies Modulators of CD47
Provisionally accepted
Ling Yin
Wei He
Yifan Wang
Huimin ZhangMin HuangYuelong YanSiting Li
Xu FengFrancisco Saenz
Jie Zhang
Dandan ZhuChang YangTiantian MaJialing Fu
Junjie Chen*- University of Texas MD Anderson Cancer Center, Houston, United States
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CD47 is a key innate immune checkpoint that enables tumor cells to evade macrophage-mediated clearance. To systematically identify genetic regulators of CD47 surface expression, we performed FACS-based genome-wide CRISPR screens in three murine cancer cell lines, B16 (melanoma), MC38 (colon adenocarcinoma), and EMT6 (breast carcinoma). Comparative analysis of cells with high or low CD47 surface expression using DrugZ revealed CD47 itself as the top hit, validating the screens. Notably, DNAJC13 emerged as a consistent and robust regulator of CD47 expression across all three cell lines. Functional validation using DNAJC13-knockout cells confirmed a significant reduction in CD47 surface levels. Furthermore, in co-culture assays with macrophages, DNAJC13-deficient tumor cells exhibited increased susceptibility to phagocytosis, supporting a functional role for DNAJC13 in innate immune evasion. Finally, we verify that DNAJC13-knockout decrease tumor burden when treated with CD47 blockade. Overall, this study highlights a previously unrecognized regulator of CD47 and demonstrates the utility of high-throughput FACS-based CRISPR screening to uncover modulators of key immune checkpoint pathways.
Keywords: FASC, CRISPR/Cas9 screen, cd47, macrophage, DnaJC13
Received: 12 Aug 2025; Accepted: 02 Dec 2025.
Copyright: © 2025 Yin, He, Wang, Zhang, Huang, Yan, Li, Feng, Saenz, Zhang, Zhu, Yang, Ma, Fu and Chen. 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: Junjie Chen
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