| 鸡胚电转染-RNA干扰技术在体模型的建立 |
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| 引用本文： | 陈大为,李瑞国,刘佳利. 鸡胚电转染-RNA干扰技术在体模型的建立[J]. 中国实验动物学报, 2010, 18(5): 390-393,I0006,I0007. DOI: 10.3969/j.issn.1005-4847.2010.05.06 |
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| 作者姓名： | 陈大为 李瑞国 刘佳利 |
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| 作者单位： | 1. 中国农业大学生物学院农业生物技术国家重点实验室,北京,100193
2. 中国生物技术发展中心,北京,100036 |
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| 摘 要： | 目的建立在体鸡胚电转染-RNA干扰技术(RNAi)模型。方法通过SOE-PCR方法,利用shRNA中的Loop环作为交叠区序列成功的建立了一种方便的shRNA表达序列构建方法,将Isl-1特异性的shRNA序列插入到pEGFP-H1-shRNA质粒中,通过注射后电转染,利用免疫组织化学方法检测Isl-1在鸡胚神经管和背根神经节(dorsal root ganglia,DRG)中的表达。结果鸡胚神经管和DRG中Isl-1的表达受到明显抑制。结论成功建立了在体鸡胚电转染-RNAi模型,为以鸡胚为模式动物研究神经管和DRG发育相关基因的功能提供了有力的工具。
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| 关 键 词： | 鸡胚 神经管 背根神经节 RNA抑制 在体电转染 |
Establishment of a Chicken Embryo Model of in Ovo RNAi-Electroporation |
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| CHEN Da-wei,LI Rui-guo,LIU Jia-li. Establishment of a Chicken Embryo Model of in Ovo RNAi-Electroporation[J]. Acta Laboratorium Animalis Scientia Sinica, 2010, 18(5): 390-393,I0006,I0007. DOI: 10.3969/j.issn.1005-4847.2010.05.06 |
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| Authors: | CHEN Da-wei LI Rui-guo LIU Jia-li |
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| Affiliation: | 1.State Key Laboratory of Agrobiotechnology,College of Biological Sciences;China Agricultural University,Beijing 100193,China;2.China National Center for Biotechnology Development,Beijing 100036) |
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| Abstract: | Objective To establish an efficient chicken embryo model of in ovo RNAi-electroporation.Methods A convenient method was established to construct the shRNA expression cassette.By this technology,the Isl-1 specific shRNA sequence was inserted into the pEGFP-H1-shRNA vector.Then the in ovo electroporation was conducted after the plasmid injection.The expression and distribution of Isl-1 were detected in the neural tube and dorsal root ganglia （DRG） by immunohistochemical staining.Results The Isl-1 expression in the neural tube and DRG of chicken embryo was significantly inhibited.Conclusion An efficient chicken embryo model of in ovo RNAi-electroporation is successfully established,and it provides a powerful tool for the studying the functions of related genes in the development of neural tube and dorsal root ganglia of chicken embryos. |
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| Keywords: | Chicken embryo Neural tube DRG RNAi in ovo electroporation |
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