鸡miR-20a靶基因TP53INP1鉴定

前期研究发现,过表达mi R-17-92基因簇能促进鸡前脂肪细胞增殖,但作用机制尚不清楚。生物信息学分析发现,TP53INP1是mi R-17-92基因簇成员mi R-17-5p和mi R-20a的潜在靶基因,研究采用荧光素酶报告基因技术和基因过表达技术开展该靶基因的验证。结果表明,过表达mi R-17-92基因簇能显著抑制TP53INP1的3＇UTR报告基因活性;而转染mi R-20a抑制剂能显著提高TP53INP1的3＇UTR报告基因活性。将mi RNA抑制剂分别转染到DF1细胞和鸡前脂肪细胞中,采用real-time RT-PCR方法检测细胞TP53INP1基因表达变化。结果显示,mi...

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Published in东北农业大学学报 Vol. 46; no. 9; pp. 69 - 74
Main Author 王宁 段逵 宋鹤 张潇飞 张天目 张文建 闫晓红 王守志 李辉
Format Journal Article
LanguageChinese
Published 农业部鸡遗传育种重点实验室,东北农业大学动物科学技术学院,哈尔滨 150030 2015
Subjects
miR-17-92基因簇
TP53INP1
前脂肪细胞
基因表达
鸡
miR-17-92基因簇
鸡
前脂肪细胞
chicken
miR-17-92 cluster
TP53INP1
基因表达
preadipocytes
gene expression
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ISSN1005-9369

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Summary:前期研究发现,过表达mi R-17-92基因簇能促进鸡前脂肪细胞增殖,但作用机制尚不清楚。生物信息学分析发现,TP53INP1是mi R-17-92基因簇成员mi R-17-5p和mi R-20a的潜在靶基因,研究采用荧光素酶报告基因技术和基因过表达技术开展该靶基因的验证。结果表明,过表达mi R-17-92基因簇能显著抑制TP53INP1的3＇UTR报告基因活性;而转染mi R-20a抑制剂能显著提高TP53INP1的3＇UTR报告基因活性。将mi RNA抑制剂分别转染到DF1细胞和鸡前脂肪细胞中,采用real-time RT-PCR方法检测细胞TP53INP1基因表达变化。结果显示,mi R-20a抑制剂能促进TP53INP1表达,TP53INP1是mi R-20a的一个靶基因。
Bibliography:23-1391/S
The previous studies showed that overexpression of miR-17-92 cluster promoted the chicken preadipocyte proliferation, however, the underlying molecular mechanism remains unclear. BioinformaUcs analysis found that TP531NP1 was a potential target gene of miR-17-5p and miR-20a encoded by the miR-17-92 cluster. In the present study, we verified this bioinformatics analysis result using a luciferase reporter assay and miRNA overexpression and knockdown. The results showed that miR-17-92 cluster significantly decreased TP531NP1 3＇ UTR luciferase reporter activity; transfection of miR-20a inhibitor significantly decreased TP531NP1 3＇ UTR luciferase reporter activity. Real-time RT-PCR expression analysis showed that transfection of miR-20a inhibitor increased the endogenous TP531NP1 expression in DFlcells and immortalized chicken preadipocytes. Taken together, these data suggest that TP531NP1 is a target gene of miR-20a.
chicken; miR-17-92 cluster; preadipocytes; TP531NP1; gene expression
WANG Ning, DUAN Kui,
ISSN:1005-9369