Preparation of a mitochondrial K+ channel Kcna 3 knockout mouse model
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KeyWord:Potassium channel, Gene-target, Embryonic stem cells, Transgenic mice, Mitochondria. Mice
张良平 同济大学附属东方医院心血管病研究室, 上海
韩俊毅 同济大学附属东方医院心血管病研究室, 上海
范慧敏 同济大学附属东方医院心血管病研究室, 上海
刘中民 同济大学附属东方医院心血管病研究室, 上海
陈炳官 同济大学附属东方医院中心实验室, 上海
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      ObjectiveKcna3 is one of mitochondrial K+ channels, which may be associated with apoptosis of myocardium during heart ischemia/reperfusion injury. The aim of this study was to establish a Kcna3 gene knockout mouse model to study the impact of mitochondrial potassium channel in heart ischemia/reperfusion injury, and explore its relationship with heart failure. MethodsA homologous recombination vector was constructed with BAC vector. Strain 129 mouse embryonic stem (ES) cells were targeted knockout of Kcna3 by the homologous recombination vector, and screened with G418 plus GANC. The Kcna3-knockout embryonic stem cells were microinjected into blastula of C57BL/6J mice after superovulation. F1 hybrid mice were bred to obtain mouse aggregation chimeras, and were identified by PCR plus sequencing of tail genomic DNA. ResultsEight Kcna3+/-mice were harvested and identified from 40 gray mice. ConclusionsThe heterozygous Kcna3-knockout mouse model is successfully established and it lays a foundation for further breed homozygous mice. It is a good model for study of the relationship between mitochondrial k+ channel abnormality with heart failure, and is of importance in research of drug screening.
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