四环素调控双链 RNA 的转录实现目的基因表达的条件性敲低。

Conditional knockdown of target gene expression by tetracycline regulated transcription of double strand RNA.

机构信息

Department of Molecular Neurobiology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.

出版信息

Dev Growth Differ. 2011 Jan;53(1):69-75. doi: 10.1111/j.1440-169X.2010.01229.x.

DOI:10.1111/j.1440-169X.2010.01229.x

Abstract

In vivo electroporation has served as an effective tool for the study of developmental biology. Here we report tetracycline inducible gene knockdown by electroporation. Our system consists of genome integration of a cassette encoding long double strand RNA (dsRNA) of a gene of interest by electroporation, transcription of which is assured by RNA polymerase II, and induction of transcription of dsRNA by tetracyclin. Long dsRNA decapped by ribozyme in the cassette and without poly A tail is processed into siRNA within nuclei. We could successfully induce knockdown of En2 and Coactosin by Dox administration.

摘要

体内电穿孔已成为发育生物学研究的有效工具。在此，我们报告了通过电穿孔实现四环素诱导的基因敲低。我们的系统包括通过电穿孔将编码目的基因的长双链 RNA（dsRNA）的盒进行基因组整合，通过 RNA 聚合酶 II 确保转录，以及通过四环素诱导 dsRNA 的转录。盒中的核酶切割 dsRNA 的帽结构并去除 poly A 尾，dsRNA 被加工成 siRNA。我们可以通过 Dox 给药成功诱导 En2 和 Coactosin 的敲低。