|
| Status |
Public on Sep 03, 2015 |
| Title |
Tracking distinct RNA populations using efficient and reversible covalent chemistry |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
We describe a chemical method to label and purify 4-thiouridine (s4U) -containing RNA. We demonstrate that methanethiolsulfonate (MTS) reagents form disulfide bonds with s4U more efficiently than the commonly used HPDP-biotin, leading to higher yields and less biased enrichment. This increase in efficiency allowed us to use s4U-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.
|
| |
|
| Overall design |
s4U metabolic labeling of RNA in 293T cells, followed by biochemical enrichment of labeled RNA with two biotinylation reagents, RNAs >200nt and miRNAs in separate experiments
|
| |
|
| Contributor(s) |
Duffy EE, Rutenberg-Shoenberg M, Stark CD, Kitchen RR, Gerstein MB, Simon MD |
| Citation(s) |
26340425 |
| Submission date |
Sep 02, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Erin Elizabeth Duffy |
| E-mail(s) |
erin_duffylacy@hms.harvard.edu
|
| Phone |
(617)432-7795
|
| Organization name |
Harvard Medical School
|
| Department |
Neurobiology
|
| Lab |
Michael E. Greenberg
|
| Street address |
200 Longwood Ave, Goldenson 405
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
|
| Samples (24)
|
|
| Relations |
| BioProject |
PRJNA260045 |
| SRA |
SRP045983 |
Less...
More...