Efficient gene knockout in primary human and murine myeloid cells by non-viral delivery of CRISPR-Cas9.

ABSTRACT: Myeloid cells play critical and diverse roles in mammalian physiology, including tissue development and repair, innate defense against pathogens, and generation of adaptive immunity. As cells that show prolonged recruitment to sites of injury or pathology, myeloid cells represent therapeutic targets for a broad range of diseases. However, few approaches have been developed for gene editing of these cell types, likely owing to their sensitivity to foreign genetic material or virus-based manipulation. Here we describe optimized strategies for gene disruption in primary myeloid cells of human and murine origin. Using nucleofection-based delivery of Cas9-ribonuclear proteins (RNPs), we achieved near population-level genetic knockout of single and multiple targets in a range of cell types witho

SUBMITTER: Freund EC 

PROVIDER: S-EPMC7336301 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature