Generation of knock-out primary and expanded human nk cells using cas9 ribonucleoproteins
AssigneeResearch Institute At Nationwide Children's Hospital
InventorsDean Anthony LEE, Meisam NAEIMI KARAROUDI
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Abstract
Disclosed are compositions and methods for genetically engineering NK cells.
Claims18
A method of genetically modifying an NK cell comprising
- a) obtaining guide RNA (gRNA) specific for the target DNA sequence;
- and
- b) introducing via electroporation into a target NK cell, a ribonucleoprotein (RNP) complex comprising a class 2 CRISPR/Cas endonuclease (Cas9) complexed with a corresponding CRISPR/Cas guide RNA that hybridizes to a target sequence within the genomic DNA of the NK cell.
A genetically modified NK cell comprising a knockout of the gene encoding the transforming growth factor-β receptor 2 (TGFBR2) or hypoxanthine phosphoribosyltransferase 1 (HPRT1).
A method of adoptively transferring an engineered NK cells to a subject in need thereof said method comprising
- a) obtaining a target NK cell to be modified;
- b) obtaining gRNA specific for the target DNA sequence;
- c) introducing via electroporation into the target NK cell, a RNP complex comprising a class 2 CRISPR/Cas endonuclease (Cas9) complexed with a corresponding CRISPR/Cas guide RNA that hybridizes to a target sequence within the genomic DNA of the target NK cell creating an engineered NK cell;
- and
- d) transferring the engineered NK cell into the subject.
The A method of treating a cancer in a subject comprising administering to the subject an NK cell that has been modified to comprise a knockout of the TGFBR2 gene.
Dependencies are inferred from the claim text.