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Updated: Nov 20, 2025
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Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
Published on: June 14, 2018
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Drug target validation in primary human natural killer cells using CRISPR RNP
Jai Rautela1,2, Elliot Surgenor1, Nicholas D Huntington1,2
1Molecular Immunology Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
Journal of Leukocyte Biology
|January 19, 2021
Summary
Researchers developed an efficient CRISPR-Cas9 gene editing method for human NK cells, overcoming viral delivery challenges. This breakthrough advances NK cell immunotherapy for cancer treatment.
Area of Science:
- Immunology
- Molecular Biology
- Cancer Research
Background:
- Chimeric antigen receptor (CAR) T cell therapy has advanced cancer immunotherapy.
- Developing NK (natural killer) cell immunotherapies is hindered by resistance to viral gene delivery.
Purpose of the Study:
- To establish an efficient gene editing technique for human NK cells.
- To validate the role of specific gene deletions in NK cell antitumor activity.
Main Methods:
- Utilized electroporation and CRISPR-Cas9 ribonucleoprotein (RNP) complexes for genetic modification of human NK cells.
- Optimized electroporation parameters and buffers for enhanced protein delivery and cell viability.
- Deleted NCR1 and CISH genes in primary human NK cells.
Main Results:
- Demonstrated an efficient method for genetically editing human NK cells via electroporation and CRISPR-Cas9 RNP.
- Validated the crucial roles of NCR1 and CISH in regulating NK cell-mediated antitumor functions, consistent with murine models.
- Showcased the potential of this technique for advancing NK cell immunotherapy drug discovery.
Conclusions:
- Electroporation with CRISPR-Cas9 RNPs provides an efficient platform for human NK cell genetic engineering.
- This method overcomes limitations of viral gene delivery for NK cell-based therapies.
- The findings support the development of novel NK cell immunotherapies for cancer treatment.
