CLONING AND FUNCTIONAL ANALYSIS OF SHEEP U6 PROMOTERS
CLONING AND FUNCTIONAL ANALYSIS OF SHEEP U6 PROMOTERS
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DOI：
10.1080/10495398.2011.580669
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发表时间：
2011-01-01
影响因子：
3.7
通讯作者：
Chen, Chuangfu
中科院分区：
文献类型：
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作者：
Hu, Shengwei;Ni, Wei;Chen, Chuangfu
小干扰RNA介导的基因沉默已成为调控基因表达的有力生物学工具。为了开发一种有效的短发夹RNA（shRNA）表达载体，特别是用于绵羊物种，我们已经确定了两个绵羊U6启动子的基础上高度保守的聚合酶III启动子元件。通过U6启动子驱动的shRNA抑制增强的绿色荧光蛋白（EGFP）表达来测量启动子活性。敲低实验表明，绵羊U6启动子和小鼠U6启动子诱导的EGFP敲低水平相似。这些结果表明，两个绵羊U6启动子可以有效地驱动shRNA表达基因沉默，并可能在基于RNAi的绵羊研究中应用。
Gene silencing mediated by small interfering RNA has become a powerful biological tool for the regulation of gene expression. In order to develop an effective short hairpin RNA (shRNA) expression vector, specifically for use in sheep species, we have identified two sheep U6 promoters based on the highly conserved polymerase III promoter elements. Promoter activity was measured by U6 promoter-driven shRNA to suppress enhanced green fluorescent protein (EGFP) expression. The knock down assay demonstrated that the two sheep U6 promoters and mouse U6 promoter induced a similar level of EGFP knockdown. These results suggest that the two sheep U6 promoters could efficiently drive shRNA expression for gene silencing and may have applications in RNAi-based sheep research.