Document Detail


Trapping cardiac recessive mutants via expression-based insertional mutagenesis screening.
MedLine Citation:
PMID:  23283723     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
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
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2013-01-02
Journal Detail:
Title:  Circulation research     Volume:  112     ISSN:  1524-4571     ISO Abbreviation:  Circ. Res.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-15     Completed Date:  2013-04-16     Revised Date:  2014-02-18    
Medline Journal Info:
Nlm Unique ID:  0047103     Medline TA:  Circ Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  606-17     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Genetically Modified
Breeding
Cardiomegaly / genetics*
DNA Transposable Elements / genetics
Embryo, Nonmammalian / metabolism,  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,  genetics
Mutagenesis, Insertional*
Organ Specificity
Phenotype
Zebrafish / embryology,  genetics*,  growth & development
Zebrafish Proteins / genetics*,  physiology
Grant Support
ID/Acronym/Agency:
DA14546/DA/NIDA NIH HHS; GM63904/GM/NIGMS NIH HHS; HG006431/HG/NHGRI NIH HHS; HL107304/HL/NHLBI NIH HHS; R01 GM063904/GM/NIGMS NIH HHS; R01 HG006431/HG/NHGRI NIH HHS; R01 HL081753/HL/NHLBI NIH HHS; R01 HL107304/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/DNA Transposable Elements; 0/Luminescent Proteins; 0/Zebrafish Proteins; 0/red fluorescent protein
Comments/Corrections

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