Researchers in the Department of Microbiology, Immunology, and Molecular Genetics at UCLA have developed an optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein gene editing of primary innate immune cells.
BACKGROUND:
CRISPR (clustered, regularly interspaced, short palindromic repeats) genome engineering has become a powerful tool to investigate the complex mechanisms of immune system regulation functionally. While decades of work have aimed to reprogram innate immunity genetically, the utility of current approaches are restricted by poor knockout efficiencies or have limited specificity for mature cell lineages in vivo. The field lacks an efficient and optimized strategy for editing primary innate immune cells for gene knockout and gene insertion.
INNOVATION:
Timothy O’Sullivan and colleagues in the Department of Microbiology, Immunology, and Molecular Genetics at UCLA have developed an optimized non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing strategy for primary innate immune cells that results in an almost complete loss of target gene expression from single electroporation. The method allows for any gene deletion in primary human innate immune cells. This gene-editing technology can be used to delete inhibitory molecules for use in adoptive cell therapy as a cancer treatment or to manipulate gene expression in innate immune cells to treat type 1 diabetes or other autoimmune diseases.
POTENTIAL APPLICATIONS:
• Nucleofection
• Diagnostics
• Gene Knockout
• Gene Knock-in
• In Vivo and in Vitro Gene Therapy
• Treatment of Type 2 Diabeties
• Adoptive Cell Transfer Therapy (Cancer)
ADVANTAGES:
• High throughput
• Cost Effective
• More efficient and optimized than current innate immune knockout strategies
DEVELOPMENT-TO-DATE: The invention has been designed and a prior proof of concept has been demonstrated in a paper published in Cell Reports in 2020. To date this method has been validated in human peripheral blood-derived monocyte derived macrophages, natural killer cells, and monocyte derived dendritic cells.
RELATED PAPERS:
Riggan, L., Hildreth, A.D., Rolot, M., Wong Y.Y., Satyadi, W., Sun, R., Huerta, C., and O’Sullivan, T.E. CRISPR Cas9 ribonucleoprotein-mediated genomic editing in primary mature innate immune cells. Cell Reports (2020). PMID: 32433960