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Master regulators and cofactors of human neuronal cell fate specification identified by CRISPR gene activation screens [CAS_TF_sgNGN3_Screen_AmpliconSeq]

ABSTRACT: Master regulators and cofactors of human neuronal cell fate specification identified by CRISPR gene activation screens [CAS_TF_sgNGN3_Screen_AmpliconSeq]

PROVIDER: PRJNA668361 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12802763_1.fastq.gz	Fastqsanger.gz
	SRR12802763_2.fastq.gz	Fastqsanger.gz
	SRR12802764_1.fastq.gz	Fastqsanger.gz
	SRR12802764_2.fastq.gz	Fastqsanger.gz
	SRR12802765_1.fastq.gz	Fastqsanger.gz

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Project description:Technologies to reprogram cell-type specification have revolutionized the fields of regenerative medicine and disease modeling. Currently, the selection of fate-determining factors for cell reprogramming applications is typically a laborious and low-throughput process. Due to the diversity of neuronal subtypes governed by a variety of distinct regulators, methods for the programmed specification of neurons would particularly benefit from a systematic understanding of the transcriptional codes that define the neuronal phenotype. Therefore, we used high-throughput pooled CRISPR activation (CRISPRa) screens to map human neuronal cell-fate regulators. We utilized dCas9-based activation to target 1,496 putative transcription factors in the human genome. Using a reporter of neuronal commitment, we profiled the neurogenic activity of these transcription factors in human pluripotent stem cells, leading to a curated set of neuronal factors. Activation of pairs of transcription factors revealed neuronal cofactors, including E2F7, RUNX3 and LHX8, that improve conversion efficiencies and influence gene expression programs related to subtype-specificity and maturation of neuronal cell types. Finally, using multiplexed gene regulation with orthogonal CRISPR systems, we demonstrated improved neuronal differentiation with concurrent activation and repression of target genes, underscoring the power of CRISPR-based gene regulation for programming complex cellular phenotypes.

2020-12-01 | GSE159340 | GEO