Efficient generation of isogenic primary human myeloid cells using CRISPR-Cas9 ribonucleoproteins.

ABSTRACT: Genome engineering of primary human cells with CRISPR-Cas9 has revolutionized experimental and therapeutic approaches to cell biology, but human myeloid-lineage cells have remained largely genetically intractable. We present a method for the delivery of CRISPR-Cas9 ribonucleoprotein (RNP) complexes by nucleofection directly into CD14⁺ human monocytes purified from peripheral blood, leading to high rates of precise gene knockout. These cells can be efficiently differentiated into monocyte-derived macrophages or dendritic cells. This process yields genetically edited cells that retain transcript and protein markers of myeloid differentiation and phagocytic function. Genetic ablation of the restriction factor SAMHD1 increased HIV-1 infection >50-fold, demonstrating the power of thi

SUBMITTER: Hiatt J 

PROVIDER: S-EPMC8188731 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature