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| Status |
Public on Mar 01, 2020 |
| Title |
OTX2 represses sister cell fate choices in the developing retina to promote photoreceptor specification |
| Organism |
Gallus gallus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
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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| Overall design |
10x-based single cell RNA sequencing from retinal cells electroporated at E5 and cultured in explant form for 48h.
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| Contributor(s) |
Ghinia Tegla MG, Emerson M |
| Citation(s) |
32347797 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 EY024982 |
Transcriptional Regulation of Cone Photoreceptor Genesis |
THE CITY COLLEGE OF THE CITY UNIVERSITY OF NEW YORK |
MARK M EMERSON |
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| Submission date |
Dec 18, 2019 |
| Last update date |
Jun 01, 2020 |
| Contact name |
Miruna Georgiana Ghinia Tegla |
| E-mail(s) |
mghinia@ccny.cuny.edu
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| Organization name |
CCNY CUNY
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| Department |
Biology
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| Lab |
Emerson Lab
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| Street address |
85 St. Nicholas Terrace
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| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10031 |
| Country |
USA |
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| Platforms (1) |
| GPL19005 |
Illumina HiSeq 2500 (Gallus gallus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA596379 |
| SRA |
SRP238072 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE142244_RAW.tar |
38.9 Mb |
(http)(custom) |
TAR (of MTX, TSV) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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