CRISPR–Cas9 Genetic Analysis of Virus–Host Interactions

• Main Authors: Makda Gebre, Jason L. Nomburg, Benjamin E. Gewurz
• Format: Article
• Language: English
• Published: MDPI AG 2018-01-01
• Series: Viruses
• Subjects: genome engineering; CRISPR screen; Cas9; Epstein–Barr virus; host dependency factor; CRISPR interference; CRISPR activation; combinatorial CRISPR
• Online Access: http://www.mdpi.com/1999-4915/10/2/55

Clustered regularly interspaced short palindromic repeats (CRISPR) has greatly expanded the ability to genetically probe virus–host interactions. CRISPR systems enable focused or systematic, genomewide studies of nearly all aspects of a virus lifecycle. Combined with its relative ease of use and high reproducibility, CRISPR is becoming an essential tool in studies of the host factors important for viral pathogenesis. Here, we review the use of CRISPR–Cas9 for the loss-of-function analysis of host dependency factors. We focus on the use of CRISPR-pooled screens for the systematic identification of host dependency factors, particularly in Epstein–Barr virus-transformed B cells. We also discuss the use of CRISPR interference (CRISPRi) and gain-of-function CRISPR activation (CRISPRa) approaches to probe virus–host interactions. Finally, we comment on the future directions enabled by combinatorial CRISPR screens.