Establishment of conditional vectors for hairpin siRNA knockdowns

被引：139

作者：

Matsukura, S ^{[1
]}

Jones, PA ^{[1
]}

Takai, D ^{[1
]}

机构：

[1] Univ So Calif, Keck Sch Med, USC Norris Comprehens Canc Ctr, Dept Biochem & Mol Biol, Los Angeles, CA 90033 USA

来源：

NUCLEIC ACIDS RESEARCH | 2003年 / 31卷 / 15期

关键词：

D O I：

10.1093/nar/gng077

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Small interference RNA (siRNA) is an emerging methodology in reverse genetics. Here we report the development of a new tetracycline-inducible vector-based siRNA system, which uses a tetracycline-responsive derivative of the U6 promoter and the tetracycline repressor for conditional in vivo transcription of short hairpin RNA. This method prevents potential lethality immediately after transfection of a vector when the targeted gene is indispensable, or the phenotype of the knockdown is lethal or results in a growth abnormality. We show that the controlled knockdown of DNA methyltransferase 1 (DNMT1) in human cancer resulted in growth arrest. Removal of the inducer, doxycycline, from treated cells led to re-expression of the targeted gene. Thus the method allows for a highly controlled approach to gene knockdown.

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页数：5

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