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张良平,韩俊毅,范慧敏,刘中民,陈炳官.线粒体钾通道Kcna3基因敲除小鼠初步制备[J].中国实验动物学报,2012,(1):34~37.
线粒体钾通道Kcna3基因敲除小鼠初步制备
Preparation of a mitochondrial K+ channel Kcna 3 knockout mouse model
  
DOI:10.3969/j.issn.1005-4847.2012.01.008
中文关键词:  钾通道  基因打靶  胚胎干细胞  转基因小鼠  线粒体
英文关键词:Potassium channel, Gene-target, Embryonic stem cells, Transgenic mice, Mitochondria. Mice
基金项目:上海市科委科研基金(项目编号:064119504);上海市浦东新区重点学科群建设项目(项目号:Pkzxkq2010-01);上海市科委基础重点科研基金(项目号:10JC1413400)资助。
作者单位E-mail
张良平 同济大学附属东方医院心血管病研究室, 上海200120 frankfan64@hotmail.com 
韩俊毅 同济大学附属东方医院心血管病研究室, 上海200120  
范慧敏 同济大学附属东方医院心血管病研究室, 上海200120  
刘中民 同济大学附属东方医院心血管病研究室, 上海200120  
陈炳官 同济大学附属东方医院中心实验室, 上海200120  
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中文摘要:
      目的为观察线粒体钾通道在缺血再灌注(I/R)心肌损伤中的作用,探讨其和心衰的关系,制备基因敲除小鼠模型以探讨钾通道单分子作用。方法用BAC载体制备同源重组载体,对129小鼠胚胎干细胞(ES)打靶筛选后,显微注射至C57BL/6J小鼠囊胚获得嵌合小鼠。经尾基因组DNA PCR鉴定和测序,鉴别杂合子小鼠。结果在40只灰色小鼠中初步鉴定出Kcna3+/-基因型F1小鼠8只。结论在国内首先用ES同源重组基因打靶方法,成功育成Kcna3基因敲除鼠杂合子,为下一步获得纯合子鼠奠定了基础。对进一步用钾离子通道病模型研究心肌保护病理生理机制和药物筛选具重要意义。
英文摘要:
      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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