PUBLICATION

Mutagenesis of putative ciliary genes with the CRISPR/Cas9 system in zebrafish identifies genes required for retinal development

Authors
Hu, R., Huang, W., Liu, J., Jin, M., Wu, Y., Li, J., Wang, J., Yu, Z., Wang, H., Cao, Y.
ID
ZDB-PUB-190110-6
Date
2019
Source
FASEB journal : official publication of the Federation of American Societies for Experimental Biology   33(4): 5248-5256 (Journal)
Registered Authors
Cao, Ying
Keywords
cilium, genetic screen, retina
MeSH Terms
  • Animals
  • CRISPR-Associated Protein 9/genetics
  • CRISPR-Associated Protein 9/metabolism*
  • Cell Cycle Proteins/genetics
  • Cell Cycle Proteins/metabolism
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/physiology*
  • In Situ Hybridization
  • Mutagenesis
  • Neural Tube/embryology
  • Neural Tube/metabolism
  • Protein Serine-Threonine Kinases/genetics
  • Protein Serine-Threonine Kinases/metabolism
  • Proto-Oncogene Proteins/genetics
  • Proto-Oncogene Proteins/metabolism
  • Retina/embryology*
  • Retina/metabolism*
  • Zebrafish
  • Zebrafish Proteins/metabolism*
PubMed
30624971 Full text @ FASEB J.
Abstract
Cilia are conserved microtubule-based organelles that function as mechanical and chemical sensors in various cell types. By bioinformatic, genomic, and proteomic studies, more than 2000 proteins have been identified as cilia-associated proteins or putative ciliary proteins; these proteins are referred to as the ciliary proteome or the ciliome. However, little is known about the function of these numerous putative ciliary proteins in cilia. To identify the possible new functional proteins or pathways in cilia, we carried out a small-scale genetic screen targeting 54 putative ciliary genes by using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system. We successfully constructed 54 zebrafish mutants, and 8 of them displayed microphthalmias. Three of these 8 genes encode proteins for protein transport, suggesting the important roles of protein transport in retinal development. In situ hybridization revealed that all these genes are expressed in zebrafish eyes. Furthermore, polo-like kinase 1 was required for ciliogenesis in neural tube. We uncovered the potential function of the ciliary genes for the retinal development of zebrafish.-Hu, R., Huang, W., Liu, J., Jin, M., Wu, Y., Li, J., Wang, J., Yu, Z., Wang, H., Cao, Y. Mutagenesis of putative ciliary genes with the CRISPR/Cas9 system in zebrafish identifies genes required for retinal development.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping