Tracking distinct RNA populations using efficient and reversible covalent chemistry

Details

• Alternative Title:
  Mol Cell
• Personal Author:
  Duffy, Erin E. ; Rutenberg-Schoenberg, Michael ; Stark, Catherine D. ; Kitchen, Robert R. ; Gerstein, Mark B. ; Simon, Matthew D.
• Description:
  We describe a chemical method to label and purify 4-thiouridine (s(4)U)-containing RNA. We demonstrate that methanethiosulfonate (MTS) reagents form disulfide bonds with s(4)U more efficiently than the commonly used HPDP-biotin, leading to higher yields and less biased enrichment. This increase in efficiency allowed us to use s(4)U labeling to study global microRNA (miRNA) turnover in proliferating cultured human cells without perturbing global miRNA levels or the miRNA processing machinery. This improved chemistry will enhance methods that depend on tracking different populations of RNA, such as 4-thiouridine tagging to study tissue-specific transcription and dynamic transcriptome analysis (DTA) to study RNA turnover.
• Subjects:
  Article Biotin Cell Proliferation Disulfides Gene Expression Profiling HEK293 Cells Humans Indicators And Reagents Mesylates MicroRNAs Organic Chemistry Processes RNA Processing, Post-Transcriptional Thiouridine

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• Source:
  Mol Cell. 59(5):858-866.
• Pubmed ID:
  26340425
• Pubmed Central ID:
  PMC4560836
• Document Type:
  Journal Article
• Funding:
  DP2 1DP2HD083992/DP/NCCDPHP CDC HHS/United States ; DP2 HD083992/HD/NICHD NIH HHS/United States ; T32 GM007223/GM/NIGMS NIH HHS/United States
• Volume:
  59
• Issue:
  5
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:5a6482c1ccdbad081c24fe613f60511cb3afde2077d1e83853eb5621f8dae97d
• Download URL:
  https://stacks.cdc.gov/view/cdc/36272/cdc_36272_DS1.pdf
• File Type:
  [PDF - 917.69 KB ]