High-throughput identification of RNA nuclear enrichment sequences. Journal Article uri icon

Overview

abstract

  • In the post-genomic era, thousands of putative noncoding regulatory regions have been identified, such as enhancers, promoters, long noncoding RNAs (lncRNAs), and a cadre of small peptides. These ever-growing catalogs require high-throughput assays to test their functionality at scale. Massively parallel reporter assays have greatly enhanced the understanding of noncoding DNA elements en masse Here, we present a massively parallel RNA assay (MPRNA) that can assay 10,000 or more RNA segments for RNA-based functionality. We applied MPRNA to identify RNA-based nuclear localization domains harbored in lncRNAs. We examined a pool of 11,969 oligos densely tiling 38 human lncRNAs that were fused to a cytosolic transcript. After cell fractionation and barcode sequencing, we identified 109 unique RNA regions that significantly enriched this cytosolic transcript in the nucleus including a cytosine-rich motif. These nuclear enrichment sequences are highly conserved and over-represented in global nuclear fractionation sequencing. Importantly, many of these regions were independently validated by single-molecule RNA fluorescence in┬ásitu hybridization. Overall, we demonstrate the utility of MPRNA for future investigation of RNA-based functionalities.

publication date

  • March 15, 2018

Full Author List

  • Shukla CJ; McCorkindale AL; Gerhardinger C; Korthauer KD; Cabili MN; Shechner DM; Irizarry RA; Maass PG; Rinn JL

Other Profiles

Additional Document Info

volume

  • 37

issue

  • 6