Identification of HIV-1 sequences targeted by the miRNA Induced silencing complex (miRISC) using Argonaute 2 cross-linking and immunoprecipitation (HITS-CLIP)

Abstract

Background The RNA interference pathway (RNAi) regulates nearly 50 percent of human transcripts at a post-transcriptional level. The effector complex of the pathway is the microRNA-induced silencing complex (miRISC) composed of one of the four Argonaute proteins (Ago1 to 4) associated with a miRNA. Once loaded into the miRISC, these small non-coding RNA guide the complex to specific mRNA sequences and the binding of the Ago protein to the transcripts can result either in its degradation or in translational repression. Several studies have shown that the miRNA pathway can also participate in the regulation of HIV-1 replication and latency. However, the molecular mechanisms involved remain to be explored. Using a HITS-CLIP (high throughput sequencing of RNA isolated by cross-linking and immunoprecipitation) approach, we identified viral RNA sequences targeted by the miRISC.