PUBLICATION
Deep Brain Photoreceptor (val-opsin) Gene Knockout Using CRISPR/Cas Affects Chorion Formation and Embryonic Hatching in the Zebrafish
- Authors
- Hang, C.Y., Moriya, S., Ogawa, S., Parhar, I.S.
- ID
- ZDB-PUB-161030-7
- Date
- 2016
- Source
- PLoS One 11: e0165535 (Journal)
- Registered Authors
- Hang, Chong-Yee, Ogawa, Satoshi
- Keywords
- Embryos, Zebrafish, Frameshift mutation, Sequence motif analysis, Chorion, Mutation detection, Phenotypes, Mutant genotypes
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Brain/embryology
- Brain/metabolism*
- CRISPR-Cas Systems/genetics*
- Chorion/growth & development*
- Female
- Gene Knockout Techniques*
- Male
- Mutation
- Opsins/chemistry
- Opsins/deficiency
- Opsins/genetics*
- Receptors, G-Protein-Coupled/chemistry
- Receptors, G-Protein-Coupled/deficiency
- Receptors, G-Protein-Coupled/genetics*
- Zebrafish/embryology*
- Zebrafish/genetics*
- PubMed
- 27792783 Full text @ PLoS One
Citation
Hang, C.Y., Moriya, S., Ogawa, S., Parhar, I.S. (2016) Deep Brain Photoreceptor (val-opsin) Gene Knockout Using CRISPR/Cas Affects Chorion Formation and Embryonic Hatching in the Zebrafish. PLoS One. 11:e0165535.
Abstract
Non-rod non-cone photopigments in the eyes and the brain can directly mediate non-visual functions of light in non-mammals. This was supported by our recent findings on vertebrate ancient long (VAL)-opsin photopigments encoded by the val-opsinA (valopa) and val-opsinB (valopb) genes in zebrafish. However, the physiological functions of valop isoforms remain unknown. Here, we generated valop-mutant zebrafish using CRISPR/Cas genome editing, and examined the phenotypes of loss-of-function mutants. F0 mosaic mutations and germline transmission were confirmed via targeted insertions and/or deletions in the valopa or valopb gene in F1 mutants. Based on in silico analysis, frameshift mutations converted VAL-opsin proteins to non-functional truncated forms with pre-mature stop codons. Most F1 eggs or embryos from F0 female valopa/b mutants showed either no or only partial chorion elevation, and the eggs or embryos died within 26 hour-post-fertilization. However, most F1 embryos from F0 male valopa mutant developed but hatched late compared to wild-type embryos, which hatched at 4 day-post-fertilization. Late-hatched F1 offspring included wild-type and mutants, indicating the parental effects of valop knockout. This study shows valop gene knockout affects chorion formation and embryonic hatching in the zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping