Identifying proximal RNA interactions from cDNA-encoded crosslinks with ShapeJumper

PLoS Comput Biol. 2021 Dec 14;17(12):e1009632. doi: 10.1371/journal.pcbi.1009632. eCollection 2021 Dec.

Abstract

SHAPE-JuMP is a concise strategy for identifying close-in-space interactions in RNA molecules. Nucleotides in close three-dimensional proximity are crosslinked with a bi-reactive reagent that covalently links the 2'-hydroxyl groups of the ribose moieties. The identities of crosslinked nucleotides are determined using an engineered reverse transcriptase that jumps across crosslinked sites, resulting in a deletion in the cDNA that is detected using massively parallel sequencing. Here we introduce ShapeJumper, a bioinformatics pipeline to process SHAPE-JuMP sequencing data and to accurately identify through-space interactions, as observed in complex JuMP datasets. ShapeJumper identifies proximal interactions with near-nucleotide resolution using an alignment strategy that is optimized to tolerate the unique non-templated reverse-transcription profile of the engineered crosslink-traversing reverse-transcriptase. JuMP-inspired strategies are now poised to replace adapter-ligation for detecting RNA-RNA interactions in most crosslinking experiments.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Algorithms
  • Binding Sites
  • Computational Biology
  • Cross-Linking Reagents
  • DNA, Complementary / chemistry*
  • DNA, Complementary / genetics
  • Genetic Engineering
  • Models, Molecular
  • Nucleic Acid Conformation
  • RNA / chemistry*
  • RNA / genetics
  • Sequence Alignment / statistics & numerical data
  • Software*

Substances

  • Cross-Linking Reagents
  • DNA, Complementary
  • RNA