心脏特异性lncRNA-uc.167转基因小鼠的构建与表型分析
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国家自然科学基金(81370278)


Construction and phenotype analysis of heart-specific lncRNA-uc.167 transgenic mice
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    摘要:

    目的:构建lncRNA-uc.167心肌组织特异性转基因小鼠并对其进行表型分析-方法:采用SfiⅠ-AscⅠ双酶切载体质粒pRP.ExSi-Isl1和目的片段uc167-pMD18-禄R-T质粒,回收连接,从而把uc.167基因克隆入心肌组织特异表达的Isl启动子的下游,构建转基因表达载体,将所得载体线性化后,通过显微注射法建立uc.167转基因小鼠,并利用特异引物PCR法鉴定转基因小鼠的基因型-通过心脏组织学和超声检查对转基因小鼠的表型进行了初步分析-结果:成功构建 uc.167转基因小鼠;超声心动图显示转基因小鼠心脏结构及相关功能指标均未见明显异常;心脏重量指数也无明显改变;HE染色提示心室大小和室壁厚度均无显著性差异,心肌细胞大小也无明显改变,且未见炎症浸润-细胞水肿等异常现象;Masson染色表明转基因阳性和同窝阴性小鼠均未出现明显的心肌纤维化改变-结论:uc.167转基因小鼠并未出现明显的心脏发育畸形,心脏结构与功能也未见明显异常-

    Abstract:

    Objective:To construct heart-specific lncRNA-uc.167 transgenic mice and analyze their phenotype. Methods:After double enzyme digestion by Sfi1 and Asc1,Vector plasmid pRP. ExSi Isl1 and purpose fragment uc167-pMD18■ Tplasmid were recycled and connected. And then, the uc.167 gene was cloned into the downstream of the Isl promoter specifically expressed in myocardial tissue to construct the transgene expression vector. Finally,we made the resulting vector linearization,established uc.167 transgenic mice by micro injection and detected the genotype by PCR. The phenotype of transgenic mice was preliminarily analyzed by cardiac histology and echocardiography. Results:We successfully constructed uc.167 transgenic mice. Echocardiography displayed that transgenic mice had no obvious abnormality in cardiac structure and function compared with the wild type mice. Heart weight index was not significantly changed both in positive and negative transgenic mice. HE staining suggested ventricular size and ventricular wall thickness had no significant difference and there were also no abnormal phenomenon including the change of cell size,inflammatory infiltration and cell edema in myocardial cell. Masson staining showed that both positive and negative transgenic mice had no significant change in myocardial fibrosis. Conclusion:Uc.167 transgenic mice did not appear obvious cardiac malformations,and cardiac structure and function were not significantly abnormal.

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陈玉梅,宋桂仙,李 杏,范 燚,吴丽洁,李 华,张奇军,钱玲梅.心脏特异性lncRNA-uc.167转基因小鼠的构建与表型分析[J].南京医科大学学报(自然科学版),2016,(2):160-165

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  • 收稿日期:2015-09-07
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  • 在线发布日期: 2016-02-29
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