Strategic self-limiting production of infectious HIV particles by CRISPR in permissive cells.

Post-translational glycosylation of the HIV-1 envelope protein involving precursor glycan trimming by mannosyl oligosaccharide glucosidase (MOGS) is critically important for morphogenesis of virions and viral entry. Strategic editing of the MOGS gene in T lymphocytes and myeloid origin cells harboring latent proviral DNA results in the production of non-infectious particles upon treatment of cells with latency reversal agents. Controlled activation of CRISPR-MOGS by rebound HIV-1 mitigates production of infectious particles that exhibit poor ability of the virus to penetrate uninfected cells. Moreover, exclusive activation of CRISPR in cells infected with HIV-1 alleviates concern for broad off-target impact of MOGS gene ablation in uninfected cells. Combination CRISPR treatment of peripheral blood lymphocytes prepared from blood of people with HIV-1 (PWH) tailored for editing the MOGS gene (CRISPR-MOGS) and proviral HIV-1 DNA (CRISPR-HIV) revealed a cooperative impact of CRISPR treatment in inhibiting the production of infectious HIV-1 particles. Our design for genetic inactivation of MOGS by CRISPR exhibits no detectable off-target effects on host cells or any deleterious impact on cell survival and proliferation. Our findings offer the development of a new combined gene editing-based cure strategy for the diminution of HIV-1 spread after cessation of antiretroviral therapy (ART) and its elimination.
Competing Interests: K.K. and R.K. from Temple University and C.M.B. and C.R.Y.C. from Children’s National Health System are named inventors on patents that cover the viral gene editing technology for MOGS that is the subject of this article. C.M.B. and C.R.Y.C. hold patent 9,885,021 related to MOGS editing of HIV-specific T cell therapies licensed by Mana Therapeutics. C.M.B. and C.R.Y.C. are scientific cofounders of Mana Therapeutics and serve on its scientific advisory board (SAB). In addition to the foregoing interests, K.K. is a co-founder, board member (observer), and chief scientific adviser and holds equity in Excision Biotherapeutics, a biotech startup that has licensed the viral gene editing technology from Temple University for commercial development and clinical trials. T.H.B. holds equity in Excision Biotherapeutics and is a member of its SAB. T.J.C. and J.G. are members of Excision Biotherapeutics, who provided advice and assisted in the design of some part of experimental quality control of the gRNAs. All other authors have no interests to disclose.
(© 2023.)