SETD4基因敲除小鼠的构建及鉴定
黄穗， 黄梦怡， 钟玙沄， 雷烨铭， 赵舒祺， 蔡军伟， 姜勇， 刘靖华
中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (2) : 202-207.
SETD4基因敲除小鼠的构建及鉴定
Establishment and Identification of SETD4gene knockout mice
目的 构建并鉴定SETD4基因敲除小鼠，为研究SETD4的生物学功能提供动物模型。 方法 将引进的SETD4flox/+小鼠与EIIa-Cre小鼠进行杂交繁殖，得到基因型为SETD4+/-.EIIa-Cre的小鼠；再与C57BL/6小鼠杂交去除Cre酶，获得杂合子SETD4+/-小鼠；该小鼠自交获得纯合子SETD4-/-小鼠。通过PCR法鉴定子代小鼠的基因型；RT-PCR、荧光定量PCR方法鉴定纯合子的SETD4基因敲除小鼠SETD4 mRNA表达情况；HE染色观察小鼠肝、肺组织的形态学变化。 结果 PCR结果表明子代小鼠的基因型符合SETD4-/-；纯合子基因敲除小鼠SETD4 mRNA水平显著低于野生型小鼠；SETD4基因敲除小鼠肝、肺组织的形态学特征与野生型小鼠相比无明显差异。 结论 本研究基于Cre/loxp系统，成功构建并鉴定了SETD4基因敲除小鼠。
Objective To study the function of SETD4， the SETD4 gene knockout homozygous mice has been established. Methods SETD4flox/+ mice and EIIa-Cre mice were interbred，the offspring of which was genotyping SETD4+/-.EIIa-Cre were crossed with C57BL/6 mice to obtain the mice with the SETD4+/- genotype，SETD4+/- heterozygous mice were inbred and then the SETD4-/- homozygous mice were gained. PCR was used to identify the genotype of the offspring，the expression of SETD4 mRNA was detected by RT-PCR and qPCR，and morphological changes of liver and lung were observed by HE staining. Result PCR results showed genotypes of the offspring of SETD4 gene knockout mice was in accordance with SETD4-/-. Compared with the wild type mice，expression of SETD4 mRNA in SETD4 gene knockout homozygous mice was significantly decreased，and morphological characteristics of liver and lung in SETD4 gene knockout homozygous mice had no significant changes. Conclusion Wehave successfully generated SETD4 gene knockout homozygous mice which can be used for study ofSETD4 function.
SETD4 / 基因敲除 / Cre/loxp系统 / KO-First技术
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国家自然科学基金项目（No. 81072425，No.81471901）；广东省自然科学基金重点项目（ 2015A030311031）
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