期刊
MOLECULAR CELL
卷 59, 期 5, 页码 858-866出版社
CELL PRESS
DOI: 10.1016/j.molcel.2015.07.023
关键词
-
资金
- NIH [DP2 1DP2HD083992]
- Searle Scholars Program
- NSF
- Yale Science Scholars
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.
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