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MRNA-seq of Phf14 KOs and wild-type controls in mESCs and differentiated EBs, NPCs and neurons

ABSTRACT: mRNA-seq of Phf14 KOs and wild-type controls in mESCs and differentiated EBs, NPCs and neurons

PROVIDER: PRJEB44877 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR5903250.fastq.gz	Fastqsanger.gz
	ERR5903251.fastq.gz	Fastqsanger.gz
	ERR5903244.fastq.gz	Fastqsanger.gz
	ERR5903245.fastq.gz	Fastqsanger.gz
	ERR5903246.fastq.gz	Fastqsanger.gz

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mRNA-seq of Phf14 KOs and wild-type controls in mESCs and differentiated EBs, NPCs and neurons

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Project description: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. HB9-GFP mESCs were differentiated into mNPCs and mMNs. Poly-A RNA was isolated from isolated populations of mESCs, mNPCs, and mMNs for RNA-sequencing and splicing analyses.

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