A virus-packageable CRISPR screen identifies host factors mediating interferon inhibition of HIV

eLife

Julie Overbaugh¹  Jolien Vermeire¹  Daryl Humes¹  Molly OhAinle¹  Louisa Helms¹  Ferdinand Roesch¹  Michael Emerman¹  Jeffrey J Delrow²  Ryan Basom² 

[1] Divisions of Human Biology and Basic Sciences, Fred Hutchinson Cancer Research Center, Washington, United States;Genomics and Bioinformatics Shared Resource, Fred Hutchinson Cancer Research Center, Seattle, United States;

关键词: CRISPR;    HIV;    interferon-stimulated genes;    restriction factor;    screen;    virus replication;   

DOI  :  10.7554/eLife.39823

来源: DOAJ

【 摘 要 】

Interferon (IFN) inhibits HIV replication by inducing antiviral effectors. To comprehensively identify IFN-induced HIV restriction factors, we assembled a CRISPR sgRNA library of Interferon Stimulated Genes (ISGs) into a modified lentiviral vector that allows for packaging of sgRNA-encoding genomes in trans into budding HIV-1 particles. We observed that knockout of Zinc Antiviral Protein (ZAP) improved the performance of the screen due to ZAP-mediated inhibition of the vector. A small panel of IFN-induced HIV restriction factors, including MxB, IFITM1, Tetherin/BST2 and TRIM5alpha together explain the inhibitory effects of IFN on the CXCR4-tropic HIV-1 strain, HIV-1LAI, in THP-1 cells. A second screen with a CCR5-tropic primary strain, HIV-1Q23.BG505, described an overlapping, but non-identical, panel of restriction factors. Further, this screen also identifies HIV dependency factors. The ability of IFN-induced restriction factors to inhibit HIV strains to replicate in human cells suggests that these human restriction factors are incompletely antagonized.Editorial note: This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review. The Reviewing Editor's assessment is that all the issues have been addressed (see decision letter).

【 授权许可】

Unknown