Fig. 1 |. Overview of primary T-cell editing using CRISPR-Cas9 RNPs. Primary CD4⁺ T cells are isolated from donor blood and activated.

crRNPs are synthesized in vitro and delivered to the activated T cells by nucleofection. These cells are expanded for molecular validation of gene editing and downstream phenotypic assays. HIV-1 virus stocks are prepared and used to infect the cell pools, which are monitored for infection by flow cytometry over several days. Genes whose editing significantly alters the percentage of infected cells relative to non-targeting controls are candidate host factors for further mechanistic studies. KO, knockout. Adapted with permission from Hultquist et al.¹⁴, Elsevier.