Trapping Cardiac Recessive Mutants via Expression-based Insertional Mutagenesis Screening

Ding, Y., Liu, W., Deng, Y., Jomok, B., Yang, J., Huang, W., Clark, K.J., Zhong, T., Lin, X., Ekker, S.C., and Xu, X.
Circulation research   112(4): 606-617 (Journal)
Registered Authors
Clark, Karl, Deng, Yun, Ding, Yonghe, Ekker, Stephen C., Huang, Wei, Jomok, Beninio Tombe, Lin, Xueying, Liu, Weibin, Xu, Xiaolei, Yang, Jingchun, Zhong, Tao P.
gene trapping, insertional mutagenesis screen, cardiac mutants, adult recessive, zebrafish, genetics, animal models, genomics, genetic heart disease
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Breeding
  • Cardiomegaly/genetics*
  • DNA Transposable Elements/genetics
  • Embryo, Nonmammalian/metabolism
  • Embryo, Nonmammalian/pathology
  • Gene Expression Profiling*
  • Gene Expression Regulation, Developmental
  • Gene Knockdown Techniques
  • Genes, Lethal
  • Genes, Recessive*
  • Genes, Reporter
  • Genetic Testing/methods*
  • Genetic Vectors/genetics
  • Genotype
  • Heart/embryology
  • Luminescent Proteins/analysis
  • Luminescent Proteins/genetics
  • Mutagenesis, Insertional*
  • Organ Specificity
  • Phenotype
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/growth & development
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/physiology
23283723 Full text @ Circ. Res.

Rationale: Mutagenesis screening is a powerful genetic tool for probing biological mechanisms underlying vertebrate development and human diseases. However, the increased colony management efforts in vertebrates impose a significant challenge for identifying genes affecting a particular organ such as the heart, especially those exhibiting adult phenotypes upon depletion.

Objective: We aim to develop a facile approach that streamlines colony management efforts via enriching cardiac mutants, which enables us to screen for adult phenotypes.

Methods and Results: The transparency of the zebrafish embryos enabled us to score 67 stable transgenic lines generated from an insertional mutagenesis screen using a transposon-based protein trapping vector. Fifteen lines with cardiac monomeric red fluorescent protein (mRFP) reporter expression were identified. We defined the molecular nature for 10 lines and bred them to homozygosity, which led to the identification of one embryonic lethal, one larval lethal, and one adult recessive mutant exhibiting cardiac hypertrophy at one year of age. Further characterization of these mutants uncovered an essential function of methionine adenosyltransferase II, alpha a (mat2aa), in cardiogenesis, an essential function of mitochondrial ribosomal protein S18B (mrps18b) in cardiac mitochondrial homeostasis, as well as a function of DnaJ (Hsp40) homolog, subfamily B, member 6b (dnajb6b) in adult cardiac hypertrophy.

Conclusions: We demonstrate that transposon-based gene trapping is an efficient approach for identifying both embryonic and adult recessive mutants with cardiac expression. The generation of a Zebrafish Insertional Cardiac (ZIC) mutant collection shall facilitate the annotation of a vertebrate cardiac genome, as well as enable heart-based adult screens.

Genes / Markers
Show all Figures
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes