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Tracking distinct RNA populations using efficient and reversible covalent chemistry

Published Date:

Sep 3 2015

Publisher's site:

http://dx.doi.org/10.1016/j.molcel.2015.07.023 New Site Icon

Source:

Mol Cell. 59(5):858-866.

[PDF-917.69 KB]

Details:

Personal Authors:

Duffy, Erin E. ; Rutenberg-Schoenberg, Michael ; Stark, Catherine D. ; Kitchen, Robert R. ; Gerstein, Mark B. ; ... More ▼

Keywords:

[+]

Pubmed ID:

26340425

Pubmed Central ID:

PMC4560836

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.

Document Type:

Journal Article

Collection(s):

CDC Public Access

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

Main Document Checksum:

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Supporting Files:

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