Trapping cardiac recessive mutants via expression-based insertional mutagenesis screening

Yonghe Ding, Weibin Liu, Yun Deng, Beninio Jomok, Jingchun Yang, Wei Huang, Karl J. Clark, Tao P. Zhong, Xueying Lin, Stephen C. Ekker, Xiaolei Xu

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

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 on 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 reporter expression were identified. We defined the molecular nature for 10 lines and bred them to homozygosity, which led to the identification of 1 embryonic lethal, 1 larval lethal, and 1 adult recessive mutant exhibiting cardiac hypertrophy at 1 year of age. Further characterization of these mutants uncovered an essential function of methionine adenosyltransferase II, α 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 mutant collection shall facilitate the annotation of a vertebrate cardiac genome, as well as enable heart-based adult screens.

Original languageEnglish (US)
Pages (from-to)606-617
Number of pages12
JournalCirculation research
Volume112
Issue number4
DOIs
StatePublished - Feb 15 2013

Keywords

  • adult recessive
  • cardiac mutants
  • gene trapping
  • insertional mutagenesis screen
  • transposon
  • zebrafish

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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