Journal Article

A functional map of HIV-host interactions in primary human T cells

Young, Janet M., Bouhaddou, Mehdi, Simons, Lacy M., Tobin, Victoria, Woo, Jonathan M., Rathore, Ujjwal, Marson, Alexander, Haas, Paige, Hultquist, Judd F., Leenay, Ryan T., Malik, Harmit S., Doudna, Jennifer A., Wojcechowskyj, Jason A., Krogan, Nevan J., May, Andrew P., Hiatt, Joseph, McGregor, Michael J., Roth, Theodore L., Haas, Kelsey M., Cavero, Devin A., Shifrut, Eric, Nguyen, Thong T.

2022

Overview

Human Immunodeficiency Virus (HIV) relies on host molecular machinery for replication. Systematic attempts to genetically or biochemically define these host factors have yielded hundreds of candidates, but few have been functionally validated in primary cells. Here, we target 426 genes previously implicated in the HIV lifecycle through protein interaction studies for CRISPR-Cas9-mediated knock-out in primary human CD4+ T cells in order to systematically assess their functional roles in HIV replication. We achieve efficient knockout (>50% of alleles) in 364 of the targeted genes and identify 86 candidate host factors that alter HIV infection. 47 of these factors validate by multiplex gene editing in independent donors, including 23 factors with restrictive activity. Both gene editing efficiencies and HIV-1 phenotypes are highly concordant among independent donors. Importantly, over half of these factors have not been previously described to play a functional role in HIV replication, providing numerous novel avenues for understanding HIV biology. These data further suggest that host-pathogen protein-protein interaction datasets offer an enriched source of candidates for functional host factor discovery and provide an improved understanding of the mechanics of HIV replication in primary T cells. Here, Hiatt et al. report the knock-out of over 400 genes in primary CD4+ T cells to assess their functional role in HIV replication, finding 86 initial candidates of which 47 are validated as HIV host factors, including 23 with restrictive activity.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13 / 49 / 60 APPLIED LIFE SCIENCES / 631/326/596/2097 / 631/326/596/2557 / 82/58 / Candidates / CD4 antigen / CD4-Positive T-Lymphocytes - metabolism / CRISPR / Gene Editing / Genes / Genetic modification / Genome editing / HIV / HIV Infections / HIV-1 - genetics / Host Microbial Interactions - genetics / Human immunodeficiency virus / Humanities and Social Sciences / Humans / Lymphocytes / Lymphocytes T / Molecular machines / multidisciplinary / Phenotypes / Protein interaction / Proteins / Replication / Science / Science (multidisciplinary) / system virology / virus-host interactions