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RNA polymerase II mediated transcription from the polymerase III promoters in short hairpin RNA expression vector.

Abstract：

:RNA polymerase III promoters of human ribonuclease P RNA component H1, human U6, and mouse U6 small nuclear RNA genes are commonly used in short hairpin RNA (shRNA) expression vectors due their precise initiation and termination sites. During transient transfection of shRNA vectors, we observed that H1 or U6 promoters also express longer transcripts enough to express several reporter genes including firefly luciferase, green fluorescent protein EGFP, and red fluorescent protein JRed. Expression of such longer transcripts was augmented by upstream RNA polymerase II enhancers and completely inhibited by downstream polyA signal sequences. Moreover, the transcription of firefly luciferase from human H1 promoter was sensitive to RNA polymerase II inhibitor alpha-amanitin. Our findings suggest that commonly used polymerase III promoters in shRNA vectors are also prone to RNA polymerase II mediated transcription, which may have negative impacts on their targeted use.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Rumi M,Ishihara S,Aziz M,Kazumori H,Ishimura N,Yuki T,Kadota C,Kadowaki Y,Kinoshita Y

doi

10.1016/j.bbrc.2005.11.037

subject

Has Abstract

pub_date

2006-01-13 00:00:00

pages

540-7

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(05)02554-4

journal_volume

339

pub_type

杂志文章

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