一种体内高效表达反义RNA、三链形成RNA及Ribozyme的新载体

A Novel Vector for Abundant Expression of Antisense RNA, Triplex-forming RNA and Ribozyme in vivo

为了在体内高效表达反义RNA、三链形成RNA及Ribozyme ,通过PCR克隆技术构建了一种全新载体 pBSKneorU 6’。该载体利用hsnRNAU6基因表达单元的特性 ,保留了U 6RNA分子 5’端维持分子稳定性的发夹和帽式结构的序列 ,缺失了其中间参与mRNA前体剪切的部分 ,取而代之的是 3个限制性内切酶位点。Hela细胞核抽提物体外转录实验证明该载体可有效地表达U6变体RNA分子。该载体携带新霉素抗性基因neor ,因而可用于真核细胞稳定转染的筛选。

For abundant expression of antisense RNA, triplex forming RNA and Ribozyme in vivo,a novel vector pBSKneo rU6' was constructed by PCR cloning This vector contains the intact hsnRNA U6 gene expression unit, yet replacing the 61 nt sequence in the middle of U6 snRNA coding region with three restriction enzyme sites Hela nuclear extract in vitro transcription experiments demonstrated that this vector can effectively express U6 mutant RNA Containing neo r at the same time, stably transfected pBSKneo rU6' can be selected easily