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Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
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JoVE Journal Genetics
Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen

Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen

DOI:
10.3791/55255-v

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11:32 min

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May 24, 2017

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, , ,
1Stem Cell and Regenerative Biology, Genome Institute of Singapore,A*STAR, 2Computational and Systems Biology, Genome Institute of Singapore,A*STAR

Chapters

  • 00:05Title
  • 01:12Treatment of HeLa Cells with Biotinylated Psoralen RNA Extraction, and Fragmentation
  • 02:25RNA Size Selection and Elution
  • 04:41Enrichment of RNA Crosslinking Regions
  • 05:57Proximity Ligation
  • 07:34Reverse Crosslinking of Biotinylated Psoralen, and Amplification of cDNA
  • 09:13The SPLASH Workflow and Representative Results
  • 10:50Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we detail the method of Sequencing of Psoralen crosslinked, Ligated, and Selected Hybrids (SPLASH), which enables genome-wide mapping of intramolecular and intermolecular RNA-RNA interactions in vivo. SPLASH can be applied to study RNA interactomes of organisms including yeast, bacteria and humans.

Tags

Keywords: RNA-RNA InteractionsSPLASHPsoralen CrosslinkingRNA InteractomeRNA GenomicsBiotinylated PsoralenUV CrosslinkingRNA IsolationGel ElectrophoresisSize Selection

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