基于锤头状核酶结构的miRNA-221海绵表达载体的构建及其性能研究
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- 英文论文题名：The Construction and Assessment of an Expression Vector of Hammerhead Ribozyme-based MiRNA Sponge
- 论文作者：陈玉琪 ; 周翔
- 英文论文作者：Yuqi Chen ; Xiang Zhou ; College of Chemistry and Molecular Sciences ; Wuhan University
- 年：2016
- 作者机构：武汉大学化学与分子科学学院;
- 论文关键词：锤头状核酶 ; microRNA海绵 ; microRNA抑制剂
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十八分会：化学生物学
- 语种：中文
- 分类号：Q78
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十八分会 ; 化学生物学
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：250k
- 原文格式：D
- 会议级别：全国
摘要
miRNA海绵是一条含若干个miRNA靶定位点的mRNA,可像海绵一样吸附miRNA,使其无法与天然靶点结合,因此它是高效的miRNA抑制剂。但由于miRNA与靶点的相互作用是只依赖种子区域(miRNA的第2-8位),因此特异性不高。而且并不清楚miRNA海绵能否降解吸附的miRNA。因此,我们首次设计了一种基于锤头状核酶技术的,即可特异性吸附又能降解miRNA的基因沉默策略。锤头状核酶是具有催化功能的RNA小分子,可降解剪切含有GUC位点RNA序列。基于这种特点,我们找到了miR-221(第10-12位点为GUC),并构建了基于锤头状核酶结构的miRNA-221海绵表达载体(Fig.1A),可在体内表达出含有4个miRNA结合位点的mRNA,希望它可以增强miRNA海绵的特异性,并降解吸附的miRNA。实验结果表明,与突变载体(核酶区域无活性)相比,我们的方法成功将肝癌细胞中miRNA-221的量抑制了10%。
MiRNA sponge is a mRNA which containing multiple binding sites to a miRNA of interest,it functions as a miRNA inhibitor by blocking miRNA like sponge.However,the specificity of miRNA sponge is poor and it doesn't clear whether this strategy could degrade miRNA.Due to these considerations,we developed the hammerhead ribozyme-based miRNA sponge strategy,which could recognize,target and degrade miRNA simultaneously.We selected miRNA-221 and constructed an expression vector of hammerhead ribozyme-based miRNA-221 sponge.Experiment data(Fig.1B) indicated this method has successfully down regulated the miRNA-221 for 10% compare with the liver cancer cells treated with the mutant inactive vector.
MiRNA sponge is a mRNA which containing multiple binding sites to a miRNA of interest,it functions as a miRNA inhibitor by blocking miRNA like sponge.However,the specificity of miRNA sponge is poor and it doesn't clear whether this strategy could degrade miRNA.Due to these considerations,we developed the hammerhead ribozyme-based miRNA sponge strategy,which could recognize,target and degrade miRNA simultaneously.We selected miRNA-221 and constructed an expression vector of hammerhead ribozyme-based miRNA-221 sponge.Experiment data(Fig.1B) indicated this method has successfully down regulated the miRNA-221 for 10% compare with the liver cancer cells treated with the mutant inactive vector.
引文
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