HIV-CRISPR: A CRISPR/Cas9 Screening Method to Identify Genes Affecting HIV Replication

Ferdinand Roesch; Molly OhAinle

doi:10.21769/BioProtoc.3614

HIV-CRISPR: A CRISPR/Cas9 Screening Method to Identify Genes Affecting HIV Replication

Bio Protoc. 2020 May 5;10(9):e3614. doi: 10.21769/BioProtoc.3614.

Authors

Ferdinand Roesch^{1

2}, Molly OhAinle¹

Affiliations

¹ Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, USA.
² Department of Virology, Institut Pasteur, Paris, France.

Abstract

Screening with CRISPR/Cas9 technology has already led to significant discoveries in the fields of cancer biology, cell biology and virology. Because of the relatively low false discovery rates and the ability to perform high-throughput, pooled approaches, it has rapidly become the assay of choice for screening studies, including whole-genome screens. Here, we describe a CRISPR screening protocol that allows for efficient screening of the entire life cycle of HIV-1 through packaging of the HIV-CRISPR lentiviral genomes by infecting HIV-1 virus in trans.

Keywords: CRISPR screen; Cofactors; HIV; HIV-CRISPR; High-Throughput Sequencing (HTS); Restriction factor.

Abstract

Grants and funding