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Transcriptional landscape of fate choices in the sensory lineages

ABSTRACT: Transcriptional landscape of fate choices in the sensory lineages

PROVIDER: PRJNA631259 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11735900.fastq.gz	Fastqsanger.gz
	SRR11736000.fastq.gz	Fastqsanger.gz
	SRR11736100.fastq.gz	Fastqsanger.gz
	SRR11736200.fastq.gz	Fastqsanger.gz
	SRR11736300.fastq.gz	Fastqsanger.gz

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Transcriptional landscape of fate choices in the sensory lineages

Project description:Somatic sensation is defined by the existence of a diversity of primary sensory neurons with unique biological features and response profiles to external and internal stimuli. However, there is no coherent picture about how this diversity of cell states is transcriptionally generated. Here, we used deep single cell analysis to resolve fate splits and molecular biasing processes during sensory neurogenesis. Our results revealed a complex series of successive and specific transcriptional changes in post-mitotic neurons that delineate hierarchical regulatory states leading to the generation of the main sensory neuron classes. In addition, our analysis identified previously undetected early gene modules expressed long before fate determination although being clearly associated with the final sensory subtypes. Overall, fate choice in sensory neurons involves initial co-activation of counteracting regulatory determinants of alternative lineages priming intermediate post-mitotic neuronal cells for a binary cell fate choice. Hence, the diversity of sensory neurons is generated through successive bi-potential intermediates in which synchronization of relevant gene modules and concurrent repression of competing fate programs precede cell fate stabilization and final commitment.

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Project description:Development of the vertebrate eye, like many developmental systems, depends on genes that are used iteratively in multiple distinct processes. The OTX2 transcription factor is one such gene, with a requirement for eye formation, photoreceptor and retinal pigment epithelium specification, among others. During the time period of retinal fate choice, OTX2 is expressed in subsets of retinal progenitor cells with restricted fate choices. However, given the multiple roles for OTX2 and limitations of conventional conditional knockout strategies, the functional significance of its expression is unknown. The present study reports the use of CRISPR/Cas9 gene editing to produce somatic mutations of OTX2 in the chick retina, and identifies similar phenotypes to those observed in human patients. In addition, single cell RNA sequencing was used to determine the functional consequences of OTX2 gene editing by CRISPR/Cas9 on the population of cells derived from OTX2-expressing retinal progenitor cells. This not only confirms that OTX2 is required for the generation of photoreceptors, but also for maintaining the proliferative potential of cells and suppressing the formation of specific retinal fates. These include specific subtypes of retinal ganglion and horizontal cells, suggesting that in this context OTX2 functions to repress sister cell fate choices. Upregulation of key transcription factors involved in the formation of these cells was observed suggesting that OTX2 is upstream of critical nodes of gene regulatory networks of these alternative fates.

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