Rapid, efficient and activation-neutral gene editing of polyclonal primary human resting CD4⁺ T cells allows complex functional analyses.

ABSTRACT: CD4⁺ T cells are central mediators of adaptive and innate immune responses and constitute a major reservoir for human immunodeficiency virus (HIV) in vivo. Detailed investigations of resting human CD4⁺ T cells have been precluded by the absence of efficient approaches for genetic manipulation limiting our understanding of HIV replication and restricting efforts to find a cure. Here we report a method for rapid, efficient, activation-neutral gene editing of resting, polyclonal human CD4⁺ T cells using optimized cell cultivation and nucleofection conditions of Cas9-guide RNA ribonucleoprotein complexes. Up to six genes, including HIV dependency and restriction factors, were knocked out individually or simultaneously and functionally characterized. Moreover, w

SUBMITTER: Albanese M 

PROVIDER: S-EPMC8748193 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature