CRISPR-Cas9 Can Inhibit HIV-1 Replication but NHEJ Repair Facilitates Virus Escape

Mol Ther. 2016 Mar;24(3):522-6. doi: 10.1038/mt.2016.24. Epub 2016 Jan 22.

Abstract

Several recent studies demonstrated that the clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9 can be used for guide RNA (gRNA)-directed, sequence-specific cleavage of HIV proviral DNA in infected cells. We here demonstrate profound inhibition of HIV-1 replication by harnessing T cells with Cas9 and antiviral gRNAs. However, the virus rapidly and consistently escaped from this inhibition. Sequencing of the HIV-1 escape variants revealed nucleotide insertions, deletions, and substitutions around the Cas9/gRNA cleavage site that are typical for DNA repair by the nonhomologous end-joining pathway. We thus demonstrate the potency of CRISPR-Cas9 as an antiviral approach, but any therapeutic strategy should consider the viral escape implications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • CRISPR-Associated Protein 9
  • CRISPR-Cas Systems*
  • Cell Line
  • DNA End-Joining Repair*
  • Endonucleases / genetics
  • Endonucleases / metabolism
  • Gene Targeting
  • Genome, Viral
  • HIV-1 / physiology*
  • Humans
  • Mutation
  • Protein Binding
  • Proviruses / genetics
  • RNA Editing
  • RNA, Guide, CRISPR-Cas Systems
  • Virus Replication*

Substances

  • Bacterial Proteins
  • RNA, Guide, CRISPR-Cas Systems
  • CRISPR-Associated Protein 9
  • Cas9 endonuclease Streptococcus pyogenes
  • Endonucleases