College of Animal Science and Technology, Shihezi University, Shihezi, China.
Anim Biotechnol. 2011 Jul-Sep;22(3):170-4. doi: 10.1080/10495398.2011.580669.
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.
小干扰 RNA 介导的基因沉默已成为调控基因表达的一种强大的生物学工具。为了开发一种有效的短发夹 RNA(shRNA)表达载体，专门用于绵羊物种，我们根据高度保守的聚合酶 III 启动子元件鉴定了两种绵羊 U6 启动子。通过 U6 启动子驱动的 shRNA 抑制增强型绿色荧光蛋白(EGFP)表达来测量启动子活性。敲低实验表明，两种绵羊 U6 启动子和小鼠 U6 启动子诱导了相似水平的 EGFP 敲低。这些结果表明，这两种绵羊 U6 启动子可以有效地驱动 shRNA 表达以实现基因沉默，并且可能在基于 RNAi 的绵羊研究中有应用。
