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CRISPR-Cas9 Ribonucleoprotein-Mediated Genomic Editing in Mature Primary Innate Immune Cells.

Abstract：

:CRISPR genome engineering has become a powerful tool to functionally investigate the complex mechanisms of immune system regulation. While decades of work have aimed to genetically reprogram innate immunity, the utility of current approaches is restricted by poor knockout efficiencies or limited specificity for mature cell lineages in vivo. Here, we describe an optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing of mature primary mouse innate lymphocyte cells (ILCs) and myeloid lineage cells that results in an almost complete loss of single or double target gene expression from a single electroporation. Furthermore, we describe in vivo adoptive transfer mouse models that can be utilized to screen for gene function during viral infection using cRNP-edited naive natural killer (NK) cells and bone-marrow-derived conventional dendritic cell precursors (cDCPs). This resource will enhance target gene discovery and offer a specific and simplified approach to gene editing in the mouse innate immune system.

journal_name

Cell Rep

journal_title

Cell reports

authors

Riggan L,Hildreth AD,Rolot M,Wong YY,Satyadi W,Sun R,Huerta C,O'Sullivan TE

doi

10.1016/j.celrep.2020.107651

subject

Has Abstract

pub_date

2020-05-19 00:00:00

pages

107651

issue

7

issn

2211-1247

pii

S2211-1247(20)30604-5

journal_volume

31

pub_type

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