Preclinical safety and biodistribution of CRISPR targeting SIV in non-human primates

Gene Ther. 2024 May;31(5-6):224-233. doi: 10.1038/s41434-023-00410-4. Epub 2023 Aug 17.


In this study, we demonstrate the safety and utility of CRISPR-Cas9 gene editing technology for in vivo editing of proviral DNA in ART-treated, virally controlled simian immunodeficiency virus (SIV) infected rhesus macaques, an established model for HIV infection. EBT-001 is an AAV9-based vector delivering SaCas9 and dual guide RNAs designed to target multiple regions of the SIV genome: the viral LTRs, and the Gag gene. The results presented here demonstrate that a single IV inoculation of EBT-001 at each of 3 dose levels (1.4 × 1012, 1.4 × 1013 and 1.4 × 1014 genome copies/kg) resulted in broad and functional biodistribution of AAV9-EBT-001 to known tissue reservoirs of SIV. No off-target effects or abnormal pathology were observed, and animals returned to their normal body weight after receiving EBT-001. Importantly, the macaques that received the 2 highest doses of EBT-001 showed improved absolute lymphocyte counts as compared to antiretroviral-treated controls. Taken together, these results demonstrate safety, biodistribution, and in vivo proviral DNA editing following IV administration of EBT-001, supporting the further development of CRISPR-based gene editing as a potential therapeutic approach for HIV in humans.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Dependovirus / genetics
  • Gene Editing* / methods
  • Genetic Therapy / methods
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Genetic Vectors / pharmacokinetics
  • Macaca mulatta*
  • Simian Acquired Immunodeficiency Syndrome* / therapy
  • Simian Acquired Immunodeficiency Syndrome* / virology
  • Simian Immunodeficiency Virus* / drug effects
  • Simian Immunodeficiency Virus* / genetics
  • Tissue Distribution