Silencing integrated SIV proviral DNA with TAR-specific CRISPR tools.

ABSTRACT:

Background

One approach for a functional HIV cure is to prevent transcription from integrated proviral DNA. A critical step in HIV transcription is the Tat protein interaction with the TAR element viral RNA. We tested the strategy of blocking this Tat-TAR interaction in the SIVmac model.

Methods

We designed five CRISPR short guiding RNAs (sgRNAs) targeting the SIVmac TAR element, along with inactive versions of Cas9 (dCas9). These sgRNA constructs were delivered as ribonucleoproteins or plasmid DNA, along with SIV DNA. The constructs were also tested in integrated viral DNA in a cell line chronically infected by SIV.

Results

The sgRNAs targeting the coding strand of the TAR element inhibited SIV RNA transcription in association with dCas9-KRAB, but not with dCas9.

SUBMITTER: Smith LM 

PROVIDER: S-EPMC7869755 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature