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Generation and validation of CRISPR-engineered human natural killer cell lines for research and therapeutic applications
- Source :
- STAR Protocols, STAR Protocols, Vol 2, Iss 4, Pp 100874-(2021)
- Publication Year :
- 2021
- Publisher :
- Elsevier, 2021.
-
Abstract
- Summary 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.<br />Graphical abstract<br />Highlights • Constitutively repressing or activating genes in human NK cells is difficult • CRISPR-engineered natural killer cells are attractive for therapeutic applications • Our approach generates stable CRISPRko, CRISPRi, and CRISPRa human NK cell lines<br />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.
- Subjects :
- Science (General)
Cell
Immunology
Cell Culture Techniques
Biology
General Biochemistry, Genetics and Molecular Biology
Natural killer cell
Cell Line
Q1-390
Genome editing
medicine
Transcriptional regulation
Protocol
CRISPR
Cytotoxic T cell
Humans
Gene
Molecular Biology
Cancer
Gene Editing
General Immunology and Microbiology
Cas9
General Neuroscience
Cell Biology
Killer Cells, Natural
medicine.anatomical_structure
Cancer research
CRISPR-Cas Systems
K562 Cells
Subjects
Details
- Language :
- English
- ISSN :
- 26661667
- Volume :
- 2
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- STAR Protocols
- Accession number :
- edsair.doi.dedup.....1833cca3a910cb968bd0387ee7983309