Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells

Nat Commun. 2015 Mar 10;6:6413. doi: 10.1038/ncomms7413.

Abstract

To combat hostile viruses, bacteria and archaea have evolved a unique antiviral defense system composed of clustered regularly interspaced short palindromic repeats (CRISPRs), together with CRISPR-associated genes (Cas). The CRISPR/Cas9 system develops an adaptive immune resistance to foreign plasmids and viruses by creating site-specific DNA double-stranded breaks (DSBs). Here we adapt the CRISPR/Cas9 system to human cells for intracellular defense against foreign DNA and viruses. Using HIV-1 infection as a model, our results demonstrate that the CRISPR/Cas9 system disrupts latently integrated viral genome and provides long-term adaptive defense against new viral infection, expression and replication in human cells. We show that engineered human-induced pluripotent stem cells stably expressing HIV-targeted CRISPR/Cas9 can be efficiently differentiated into HIV reservoir cell types and maintain their resistance to HIV-1 challenge. These results unveil the potential of the CRISPR/Cas9 system as a new therapeutic strategy against viral infections.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Western
  • CD4-Positive T-Lymphocytes / immunology*
  • CRISPR-Cas Systems / immunology*
  • DNA Primers / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Dosage / genetics
  • HEK293 Cells
  • HIV Infections / prevention & control*
  • Humans
  • INDEL Mutation / genetics
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Plasmids / genetics
  • Pluripotent Stem Cells / immunology*
  • Pluripotent Stem Cells / metabolism
  • Proviruses / genetics
  • Proviruses / metabolism

Substances

  • DNA Primers