Application of CRISPR/Cas9-Based Gene Editing in HIV-1/AIDS Therapy

Abstract

Despite the fact that great efforts have been made in the prevention and therapy of HIV-1 infection, HIV-1/AIDS remains a major threat to global human health. Highly active antiretroviral therapy (HAART) can suppress virus replication, but it cannot eradicate latent viral reservoirs in HIV-1/AIDS patients. Recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system has been engineered as an effective gene-editing technology with the potential to treat HIV-1/AIDS. It can be used to target cellular co-factors or HIV-1 genome to reduce HIV-1 infection and clear the provirus, as well as to induce transcriptional activation of latent virus in latent viral reservoirs for elimination. This versatile gene editing technology has been successfully applied to HIV-1/AIDS prevention and reduction in human cells and animal models. Here, we update the rapid progress of CRISPR/Cas9-based HIV-1/AIDS therapy research in recent years and discuss the limitations and future perspectives of its application.

Lessons Learned From Failures and Success Stories of HIV Breakthroughs: Are We Getting Closer to an HIV Cure?

“…Thus, using a combinatorial CRISPR/Cas9 with dual gRNA can counter these effects. Currently, there are five CRISPR/Cas9 approaches that can be utilized against HIV infection (Saayman et al, 2015;Xiao et al, 2019): (i) targeting the pre-integrated proviral dsDNA directly, (ii) knocking out early-stage host co-receptor factor CCR5, (iii) cutting of the viral genome for LTR, (iv) cutting of viral genes responsible for release, and (v) targeting reactivation of latent cells, followed by subsequent 'shock-and-kill' strategies using ART and antiviral mechanisms.…”

Reduce and Control: A Combinatorial Strategy for Achieving Sustained HIV Remissions in the Absence of Antiretroviral Therapy

“…The recent discovery of clustered regularly interspaced short palindromic repeats or CRISPR has revolutionized gene editing in mammalian cells. CRISPR repetitive sequences were initially discovered in bacteria [214], where they form part of a larger protective response aimed at bacteriophages and other intrusive DNAs [215,216]. Bacterial helicases such as Cas9 bind CRISPR RNAs (crRNA), which then target pathogenic DNA leading to Cas9-mediated cleavage and degradation of the external threat [216].…”

“…CRISPR repetitive sequences were initially discovered in bacteria [214], where they form part of a larger protective response aimed at bacteriophages and other intrusive DNAs [215,216]. Bacterial helicases such as Cas9 bind CRISPR RNAs (crRNA), which then target pathogenic DNA leading to Cas9-mediated cleavage and degradation of the external threat [216]. The ability of CRISPR to direct double-strand DNA cleavage by Cas9 at specific sites through the action of crRNAs (or small guide RNAs, sgRNAs) has created an immensely powerful gene editing tool.…”

“…This constraint will likely limit serial dosing of cocktails containing Cas9/dCas9. CRISPR/Cas9 technology was first applied to HIV-1 in 2013 [219], with several rapidly ensuing studies (reviewed in [216]). Proviral cleavage by CRISPR was often tested in cell lines, like HEK 293T cells and HeLa [219][220][221][222][223], which unfortunately do not resemble natural targets of HIV.…”

AIDS tedavisi için gen düzenleme çalışmaları

“…As mentioned HIV-1 strain enters the cells after it has bound to receptor of CD4 and a co-receptor like CCR5 or CXCR4. CD4 however, is a key part of the functional human system so disrupting it is not a good possible strategy so instead, the aforementioned CCR5 disruptions that were achieved via ZFNs to introduce homozygous CCR5Δ32 mutation on their CD4 cells is aimed with CRIPSR-Cas9 methods (Xiao, et al 2019). Via transfection of sgRNAs and Cas9, human embryonic kidney (HEK) 293T cells' CCR5 genes were silenced with SpCas9 by Cho et al (2013), proving CRISPR to be useful but only at 13% efficiency (Cho, et al 2013).…”