Functional screening of guide RNAs targeting the regulatory and structural HIV-1 viral genome for a cure of AIDS.

ABSTRACT:

Objective

There is an urgent need for the development of HIV-1 genome eradication strategies that lead to a permanent cure for HIV-1/AIDS. We previously reported that four guide RNAs (gRNAs) targeting HIV-1 long terminal repeats (LTR) effectively eradicated the entire HIV-1 genome. In this study, we sought to identify the best gRNAs targeting HIV-1 LTR and viral structural region and optimize gRNA pairing that can efficiently eradicate the HIV-1 genome.

Design

Highly specific gRNAs were designed using bioinformatics tools, and their capacity of guiding CRISPR-associated system 9 to cleave HIV-1 proviral DNA was evaluated using high-throughput HIV-1 luciferase reporter assay and rapid Direct-PCR genotyping.

Methods

The target seed sequences for each gRNA were cloned in

SUBMITTER: Yin C 

PROVIDER: S-EPMC4851589 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature