TALENs-mediated homozygous CCR5Δ32 mutations endow CD4+ U87 cells with resistance against HIV‑1 infection

Mol Med Rep. 2018 Jan;17(1):243-249. doi: 10.3892/mmr.2017.7889. Epub 2017 Oct 26.

Abstract

Since evidence suggests that transplantation of bone marrow stem cells with the C‑C chemokine receptor type 5 (CCR5)Δ32/Δ32 genotype may cure patients infected with human immunodeficiency virus (HIV)‑1, the present study aimed to reproduce the CCR5Δ32 mutation in cluster of differentiation (CD)4+ U87 cells using genome engineering methods. A modified transcription activator‑like effector nucleases (TALENs) technique, combined with homologous recombination for site‑specific, size‑controlled and homozygous DNA deletions, was used to reproduce the homozygous CCR5Δ32 mutation in CD4+ U87 cells. The results indicated that the frequency of the TALENs‑targeted mutation reached 50.4% without any selection, whereas homologous recombination from CCR5 to CCR5Δ32 occurred in 8.8% of targeted cells. Notably, a HIV‑1 challenge test demonstrated that CCR5Δ32/Δ32 CD4+ U87 cells were resistant to HIV infection. In conclusion, engineered CCR5Δ32/Δ32 mutations endowed CD4+ U87 cells with resistance against HIV‑1 infection; this site‑specific, size‑controlled and homozygous DNA deletion technique was able to induce precise genomic editing, i.e., the deletion or insertion of a predetermined length of DNA sequence at a specific locus throughout the genome.

MeSH terms

  • Base Sequence
  • Binding Sites
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • Cell Line
  • Disease Resistance / genetics*
  • Disease Resistance / immunology
  • HIV Infections / genetics*
  • HIV Infections / immunology
  • HIV Infections / virology
  • HIV-1* / immunology
  • Homozygote*
  • Humans
  • Models, Biological
  • Protein Binding
  • Receptors, CCR5 / genetics*
  • Sequence Deletion*
  • Transcription Activator-Like Effector Nucleases / metabolism*

Substances

  • CCR5 protein, human
  • Receptors, CCR5
  • Transcription Activator-Like Effector Nucleases