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| Status |
Public on Jan 01, 2016 |
| Title |
Splicing analyses of 46C mNPCs following PTBP depletion |
| Organism |
Mus musculus |
| Experiment type |
Other
|
| Summary |
PTBP1 and PTBP2 control alternative splicing programs during neuronal development, but the cellular functions of most PTBP1/2-regulated isoforms remain unknown. We show that PTBP1 guides developmental gene expression by regulating the transcription factor Pbx1. We identify exons that are differentially spliced when mouse embryonic stem cells (ESCs) differentiate into neuronal progenitor cells (NPCs) and neurons, and transition from PTBP1 to PTBP2 expression. We define those exons controlled by PTBP1 in ESCs and NPCs by RNA-seq analysis after PTBP1 depletion and PTBP1 crosslinking-immunoprecipitation. We find that PTBP1 represses Pbx1 exon 7 and the expression of its neuronal isoform Pbx1a in ESC. Using CRISPR-Cas9 to delete regulatory elements for exon 7, we induce Pbx1a expression in ESCs, finding that this activates transcription of specific neuronal genes including known Pbx1 targets. Thus PTBP1 controls the activity of Pbx1 and suppresses its neuronal transcriptional program prior to differentiation.
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| Overall design |
46C mESCs were differentiated in mNPCs. The mNPCs were treated with 10 nM control, Ptbp1, Ptbp2, or Ptbp1 and Ptbp2 siRNAs for 48 hours. The knockdowns were performed using 2 independent sets of siRNAs. Poly-A RNA was isolated for RNA-sequencing and splicing analyses.
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| Contributor(s) |
Linares AJ |
| Citation(s) |
26705333 |
| |
| Submission date |
Jul 16, 2015 |
| Last update date |
May 15, 2019 |
| Contact name |
Chia-ho Lin |
| E-mail(s) |
[email protected]
|
| Organization name |
UCLA
|
| Lab |
BLACK LAB
|
| Street address |
6577 MRL. 675 CHARLES E YOUNG DR. SOUTH
|
| City |
LOS ANGELES |
| State/province |
CA |
| ZIP/Postal code |
90095 |
| Country |
USA |
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| Platforms (1) |
| GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
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| Samples (8)
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| This SubSeries is part of SuperSeries: |
| GSE71179 |
The splicing regulator PTBP1 controls the activity of the transcription factor Pbx1 during neuronal differentiation |
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| Relations |
| BioProject |
PRJNA290074 |
| SRA |
SRP061227 |
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