Related Experiment Video
Updated: Apr 17, 2026
Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
Genome-wide CRISPR screens in primary human natural killer cells identify countermeasures against immunosuppressive
Quoc Viet Nguyen1, Yi-Jun Lan1, Jason Cheng-Yu Chang1
1Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Abstract:
Natural killer (NK) cells are promising effectors for cancer immunotherapy, but their efficacy is limited by immunosuppressive tumor microenvironments. To uncover strategies for enhancing NK cell function, we establish a CRISPR loss-of-function screening platform for primary human NK cells by combining BaEVRless-pseudotyped lentiviral transduction of sgRNA libraries with Cas9 protein electroporation. This platform enables genome-scale interrogation of gene function in non-transformed NK cells. Kinome-focused and genome-wide screens identify key regulators of NK cell proliferation, cytotoxicity, and resistance to prostaglandin E2 (PGE2)-mediated suppression. STK17B deletion enhances NK cell expansion, while loss of CCDC53 boosts degranulation and cytotoxicity. We also uncover the CRL5 complex-including RNF7, UBE2F, and CISH-as critical inhibitors of IL-2 signaling and effector function under PGE2 stress. These findings establish a scalable platform for CRISPR-based functional genomics in primary NK cells and reveal engineering targets to enhance NK cell persistence and efficacy in tumor microenvironments.
Insights
Researchers developed a CRISPR screening platform for natural killer (NK) cells to enhance cancer immunotherapy. This tool identified genes that improve NK cell function and resistance to tumor microenvironments.
Area of Science:
- Immunology
- Genetics
- Cancer Research
Background:
- Natural killer (NK) cells are crucial for cancer immunotherapy.
- Their effectiveness is hindered by immunosuppressive tumor microenvironments.
- Strategies to enhance NK cell function are needed.
Purpose of the Study:
- To establish a CRISPR loss-of-function screening platform for primary human NK cells.
- To identify genes regulating NK cell function, proliferation, cytotoxicity, and resistance to prostaglandin E2 (PGE2).
- To uncover targets for improving NK cell-based cancer immunotherapy.
Main Methods:
- Developed a CRISPR screening platform using lentiviral transduction of sgRNA libraries and Cas9 protein electroporation.
- Performed kinome-focused and genome-wide screens on primary human NK cells.
- Utilized screening to interrogate gene function and identify regulators of NK cell activity.
Main Results:
- Deletion of STK17B enhanced NK cell expansion.
- Loss of CCDC53 improved NK cell degranulation and cytotoxicity.
- Identified the CRL5 complex (RNF7, UBE2F, CISH) as inhibitors of IL-2 signaling and function under PGE2 stress.
Conclusions:
- Established a scalable platform for CRISPR-based functional genomics in primary NK cells.
- Identified STK17B, CCDC53, and the CRL5 complex as key regulators of NK cell function.
- Revealed potential engineering targets to enhance NK cell persistence and efficacy in tumor microenvironments for improved cancer immunotherapy.
