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。
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.
出处
《中国实验动物学报》
CAS
CSCD
2010年第5期390-393,I0006,I0007,共6页
Acta Laboratorium Animalis Scientia Sinica
