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Status |
Public on Nov 08, 2023 |
Title |
Missense mutations in CRX homeodomain cause dominant retinopathies through two distinct mechanisms [Spec-seq] |
Organism |
synthetic construct |
Experiment type |
Other
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Summary |
Homeodomain transcription factors (HD TFs) are instrumental to vertebrate development. Mutations in HD TFs have been linked to human diseases, but their pathogenic mechanisms remain elusive. Here we use Cone-Rod Homeobox (CRX) as a model to decipher the disease-causing mechanisms of two HD mutations, p.E80A and p.K88N, that produce severe dominant retinopathies. Through integrated analysis of molecular and functional evidence in vitro and in knock-in mouse models, we uncover two novel gain-of-function mechanisms: p.E80A increases transactivation of canonical CRX target genes in developing photoreceptors; p.K88N alters CRX DNA-binding specificity resulting in binding at ectopic sites and severe perturbation of CRX target gene expression. Both mechanisms produce novel retinal morphological defects and hinders photoreceptor maturation distinct from loss-of-function models. This study reveals the distinct roles of E80 and K88 residues in CRX HD regulatory functions and emphasizes the importance of transcriptional precision in normal development.
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Overall design |
Two Spec-seq libararies containing monomeric HD motif variants on the sense or antisense strand were designed to profile the binding specificity of WT and mutant CRX HDs. The two libraries were mixed in equal molar ratio in the Spec-seq experiments.
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Contributor(s) |
Zheng Y |
Citation(s) |
37963072 |
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Submission date |
Jan 24, 2023 |
Last update date |
Dec 06, 2023 |
Contact name |
Shiming Chen |
E-mail(s) |
chenshiming@wustl.edu
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Phone |
314-747-4351
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Organization name |
Washington University in St Louis
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Department |
Department of Ophthalmology & Visual Sciences
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Lab |
Shiming Chen, Ph.D.
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Street address |
517 South Euclid Avenue
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City |
Saint Louis |
State/province |
Missouri |
ZIP/Postal code |
63110 |
Country |
USA |
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Platforms (1) |
GPL17769 |
Illumina MiSeq (synthetic construct) |
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Samples (16)
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This SubSeries is part of SuperSeries: |
GSE223659 |
Missense mutations in CRX homeodomain cause dominant retinopathies through two distinct mechanisms |
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Relations |
BioProject |
PRJNA927050 |