CRISPR-Cas13 for HIV-1 RNA Targeting: Advances, Challenges, and NGS-Driven Insights

Abstract

The main obstacle to eliminating HIV-1 from the body exists because HIV-1 latent reservoirs continue to survive despite antiretroviral treatments which successfully manage HIV-1 infection. The DNA-editing method CRISPR-Cas9 faces hazards of unintended DNA changes but scientists developed CRISPR-Cas13 as a synthetic RNA-targeting nuclease that destroys viral RNA without affecting host DNA. The review presents recent scientific developments in Cas13-based HIV-1 control while showing how directing treatment at viral RNA transcripts produces better treatment results than direct treatment of proviral DNA. We study how Cas13 orthologs including Cas13a, Cas13b, Cas13d, and the compact Cas13X operate at the molecular level while we analyze their RNase activities which create potential safety risks for medical treatment. The existence of several major obstacles still needs resolution because the system experiences problems with delivering treatments to tissue reservoirs and the virus develops escape mutations and the system deals with unintentionally destroying RNA and the body responds to Cas proteins which come from bacteria. Next-generation sequencing has become an essential method which researchers use to determine target effectiveness while they track how mutations develop and study the impacts of various RNA sequences on gene expression. We proposed that the implementation of NGS-enabled personalized Cas13 treatment, which uses patient proviral sequencing to create crRNA designs and monitor post-treatment results, provides a practical solution to achieve successful clinical implementation.