PUBLICATION
A CRISPR-Cas9 mutation in sox9b long intergenic noncoding RNA (slincR) affects zebrafish development, behavior, and regeneration
- Authors
- Dasgupta, S., La Du, J.K., Garcia, G.R., Li, S., Tomono-Duval, K., Rericha, Y., Huang, L., Tanguay, R.L.
- ID
- ZDB-PUB-230525-32
- Date
- 2023
- Source
- Toxicological sciences : an official journal of the Society of Toxicology 194(2): 153-166 (Journal)
- Registered Authors
- La Du, Jane K., Tanguay, Robyn L.
- Keywords
- CRISPR-Cas9, aryl hydrocarbon receptor, lncRNA, non-coding RNA, regeneration, zebrafish
- Datasets
- GEO:GSE222975
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems
- Mutation
- Polychlorinated Dibenzodioxins*
- RNA, Long Noncoding*/genetics
- RNA, Long Noncoding*/metabolism
- RNA, Messenger/metabolism
- Receptors, Aryl Hydrocarbon/metabolism
- Regeneration
- Zebrafish
- Zebrafish Proteins/metabolism
- PubMed
- 37220911 Full text @ Toxicol. Sci.
Citation
Dasgupta, S., La Du, J.K., Garcia, G.R., Li, S., Tomono-Duval, K., Rericha, Y., Huang, L., Tanguay, R.L. (2023) A CRISPR-Cas9 mutation in sox9b long intergenic noncoding RNA (slincR) affects zebrafish development, behavior, and regeneration. Toxicological sciences : an official journal of the Society of Toxicology. 194(2):153-166.
Abstract
The role of long non-coding RNAs (lncRNAs) regulators of toxicological responses to environmental chemicals is gaining prominence. Previously, our laboratory discovered a lncRNA, sox9b long intergenic noncoding RNA (slincR), that is activated by multiple ligands of aryl hydrocarbon receptor (AHR). Within this study, we designed a CRISPR-Cas9-mediated slincR zebrafish mutant line to better understand its biological function in presence or absence of a model AHR ligand, 2,3,7,8 -Tetrachlorodibenzo-p-dioxin (TCDD). The slincRosu3 line contains an 18 bp insertion within the slincR sequence that changes its predicted mRNA secondary structure. Toxicological profiling showed that slincRosu3 is equally or more sensitive to TCDD for morphological and behavioral phenotypes. Embryonic mRNA-sequencing showed differential responses of 499 or 908 genes in slincRosu3 in absence or presence of TCDD Specifically, unexposed slincRosu3 embryos showed disruptions in metabolic pathways, suggesting an endogenous role for slincR. slincRosu3 embryos also had repressed mRNA levels of sox9b- a transcription factor that slincR is known to negatively regulate. Hence, we studied cartilage development and regenerative capacity- both processes partially regulated by sox9b. Cartilage development was disrupted in slincRosu3 embryos both in presence and absence of TCDD. slincRosu3 embryos also displayed a lack of regenerative capacity of amputated tail fins, accompanied by a lack of cell proliferation. In summary, using a novel slincR mutant line, we show that a mutation in slincR can have widespread impacts on gene expression and structural development endogenously and limited, but significant impacts in presence of AHR induction that further highlights its importance in the developmental process.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping