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Updated: Oct 14, 2025
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
Published on: June 14, 2018
Generation and validation of CRISPR-engineered human natural killer cell lines for research and therapeutic
Anil Kumar1, Sung June Lee1, Qiao Liu1
1Department of Systems Biology, Beckman Research Institute of City of Hope, Monrovia, CA 91016, USA.
Abstract:
Cytotoxic natural killer cells kill tumors and infected cells. We carried out CRISPR-based gene editing and transcriptional regulation in hard-to-manipulate NK-92 cells. NK-92-based therapies were found to be safe and efficacious in preclinical studies of cancers. Here, we have pioneered the generation and validation of NK-92 cells constitutively expressing Cas9 or dCas9 for knockout (CRISPRko), transcriptional activation (CRISPRa), or transcriptional repression (CRISPRi) of genes. Our CRISPR-engineered NK-92 cell platforms can be modified for research and off-the-shelf therapeutic applications.
Insights
We developed CRISPR-engineered NK-92 cells for cancer therapy. These cells can be modified for gene knockout, activation, or repression, offering a versatile platform for research and potential off-the-shelf treatments.
Area of Science:
- Immunology
- Cell Biology
- Gene Editing
Background:
- Natural killer (NK) cells are cytotoxic lymphocytes crucial for tumor and pathogen surveillance.
- NK-92 cells are a well-characterized human NK cell line with therapeutic potential.
- Genetic manipulation of NK-92 cells has been challenging, limiting their application.
Purpose of the Study:
- To establish a robust CRISPR-based gene editing and transcriptional regulation system in NK-92 cells.
- To create NK-92 cell platforms for gene knockout (CRISPRko), activation (CRISPRa), and repression (CRISPRi).
- To validate the utility of these engineered NK-92 cells for research and therapeutic development.
Main Methods:
- Utilized CRISPR-Cas9 and CRISPR-dCas9 systems for gene editing.
- Developed NK-92 cell lines constitutively expressing Cas9 or dCas9.
- Validated CRISPRko, CRISPRa, and CRISPRi functionalities within the NK-92 cell line.
Main Results:
- Successfully generated NK-92 cells engineered for constitutive Cas9 or dCas9 expression.
- Demonstrated the efficacy of CRISPRko, CRISPRa, and CRISPRi in modulating target genes within NK-92 cells.
- Preclinical studies showed NK-92-based therapies are safe and effective against cancers.
Conclusions:
- Pioneered CRISPR-engineered NK-92 cell platforms for versatile gene manipulation.
- These platforms offer significant potential for advancing NK cell-based cancer immunotherapy research.
- The engineered NK-92 cells provide a foundation for developing off-the-shelf therapeutic applications.
