PUBLICATION

A novel inducible mutagenesis screen enables to isolate and clone both embryonic and adult zebrafish mutants

Authors
Ma, Z., Zhu, P., Pang, M., Guo, L., Chang, N., Zheng, J., Zhu, X., Gao, C., Huang, H., Cui, Z., Xiong, J.W., Peng, J., Chen, J.
ID
ZDB-PUB-170906-6
Date
2017
Source
Scientific Reports   7: 10381 (Journal)
Registered Authors
Chen, Jun, Cui, Zongbin, Huang, Honghui, Peng, Jinrong, Xiong, Jing-Wei
Keywords
Developmental biology, Genetic techniques
MeSH Terms
  • Animals
  • Cloning, Molecular
  • Gene Expression Regulation, Developmental
  • Gene Transfer Techniques
  • Mutagenesis*
  • Phenotype
  • Transcriptional Activation
  • Zebrafish/genetics
  • Zebrafish/growth & development*
  • Zebrafish Proteins/genetics*
PubMed
28871129 Full text @ Sci. Rep.
Abstract
Conventional genetic screens for recessive mutants are inadequate for studying biological processes in the adult vertebrate due to embryonic lethality. Here, we report that a novel inducible mutagenesis system enables to study gene function in both embryonic and adult zebrafish. This system yields genetic mutants with conditional ectopic over- or under-expression of genes in F1 heterozygotes by utilizing inducible Tet-On transcriptional activation of sense or anti-sense transcripts from entrapped genes by Tol2 transposase-meditated transgenesis. Pilot screens identified 37 phenotypic mutants displaying embryonic defects (34 lines), adult fin regeneration defects (7 lines), or defects at both stages (4 lines). Combination of various techniques (such as: generating a new mutant allele, injecting gene specific morpholino or mRNA etc) confirms that Dox-induced embryonic abnormalities in 10 mutants are due to dysfunction of entrapped genes; and that Dox-induced under-expression of 6 genes causes abnormal adult fin regeneration. Together, this work presents a powerful mutagenesis system for genetic analysis from zebrafish embryos to adults in particular and other model organisms in general.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes