|
| Status |
Public on Mar 13, 2014 |
| Title |
Argonaute-bound small RNAs from promoter-proximal RNA Polymerase II |
| Organism |
Mus musculus |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
Argonaute (Ago) proteins mediate post-transcriptional gene repression by binding guide microRNAs (miRNAs) to regulate targeted RNAs. To confidently assess Agobound small RNAs, we adapted a mouse embryonic stem cell system to express a single inducible epitope-tagged Ago protein. Here, we report the small RNA profile of Agodeficient cells and determine Ago-dependent stability is a common feature of mammalian miRNAs. Considering both in vivo Ago-dependence for stability and Ago2 binding as defined by immunopurification, we have identified a novel class of non-canonical miRNAs derived from protein-coding gene promoters, which we name transcriptional start site miRNAs (TSS-miRNAs). A subset of promoter-proximal RNA polymerase II complexes produce hairpin RNAs that are processed in a DGCR8/Drosha-independent, but Dicer-dependent manner. TSS-miRNA activity is detectable endogenously, upon transfection of a mimic or by mRNA overexpression. Finally, we present evidence of differential expression and conservation in humans, suggesting important roles in gene regulation.
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| |
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| Overall design |
Examination of Ago immunoprecipitations and mESC without Ago proteins
|
| |
|
| Contributor(s) |
Zamudio JR |
| Citation(s) |
24581493 |
| |
| Submission date |
Sep 04, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Jesse R Zamudio |
| E-mail(s) |
jesse.zamudio@ucla.edu
|
| Organization name |
University of California, Los Angeles
|
| Department |
MCDB
|
| Lab |
Zamudio
|
| Street address |
610 CHARLES E. YOUNG DRIVE EAST BOX 957239
|
| City |
LOS ANGELES |
| State/province |
CA |
| ZIP/Postal code |
90095-7239 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
|
| Samples (12)
|
|
| Relations |
| BioProject |
PRJNA218026 |
| SRA |
SRP029585 |
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