Improvements to the HITS-CLIP protocol eliminate widespread mispriming artifacts.

ABSTRACT: High-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP) allows for high resolution, genome-wide mapping of RNA-binding proteins. We found that substantial mispriming during reverse transcription results in the overrepresentation of sequences complementary to the primer used for reverse transcription. Up to 45% of peaks in publicly available HITS-CLIP libraries are attributable to this artifact, and the majority of libraries have detectable levels of mispriming. We also found that standard techniques for validating miRNA-target interactions fail to differentiate between artifactual peaks and physiologically relevant peaks. Here, we present a modification to the HITS-CLIP protocol that effectively eliminates this artifact.

ORGANISM(S): Homo sapiens

PROVIDER: GSE78059 | GEO | 2016/02/19

SECONDARY ACCESSION(S): PRJNA312501

REPOSITORIES: GEO