Key result
Non-viral CRISPR-Cas9 editing achieves high-efficiency target gene expression in primary innate lymphocytes and myeloid cells.
Why the study?
Current approaches to genetically reprogram innate immunity are inefficient or nonspecific, limiting their functional investigation.
This protocol provides an optimized strategy for efficient non-viral CRISPR-Cas9 genomic editing in primary innate immune cells.
Answers are grounded in this paper and cite their sources.
May advance preclinical genetic studies of innate immunity; leaves open translation to human cells or therapy.
CRISPR-Cas9 genome engineering can be used to functionally investigate the complex mechanisms of immune system regulation. Decades of work have aimed to genetically reprogram innate immunity, but current approaches are inefficient or nonspecific, limiting their use. Here, we detail an optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing of primary innate lymphocytes (ILCs) and myeloid lineage cells, resulting in high-efficiency editing of target gene expression from a single electroporation. For complete details on the use and execution of this protocol, please refer to Riggan et al. (2020).
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Hildreth et al. (2020) studied this question. non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing was evaluated. An optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein genomic editing of primary innate lymphocytes and myeloid lineage cells resulted in high-efficiency editing of target gene expression.
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