A versatile approach to multiple gene RNA interference using microRNA-based short hairpin RNAs

被引：54

作者：

Zhu, Xiaocui ^{[1
]}

Santa, Leah A. ^{[1
]}

Chang, Mi Sook ^{[1
]}

Liu, Jamie ^{[1
]}

Zavzavadjian, Joelle R. ^{[1
]}

Wall, Estelle A. ^{[1
]}

Kivork, Christine ^{[1
]}

Simon, Melvin I. ^{[1
]}

Fraser, Iain Dc ^{[1
]}

机构：

[1] CALTECH, Div Biol, Pasadena, CA 91125 USA

来源：

BMC MOLECULAR BIOLOGY | 2007年 / 8卷

关键词：

D O I：

10.1186/1471-2199-8-98

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：

Background: Effective and stable knockdown of multiple gene targets by RNA interference is often necessary to overcome isoform redundancy, but it remains a technical challenge when working with intractable cell systems. Results: We have developed a flexible platform using RNA polymerase II promoter-driven expression of microRNA-like short hairpin RNAs which permits robust depletion of multiple target genes from a single transcript. Recombination-based subcloning permits expression of multi-shRNA transcripts from a comprehensive range of plasmid or viral vectors. Retroviral delivery of transcripts targeting isoforms of cAMP-dependent protein kinase in the RAW264.7 murine macrophage cell line emphasizes the utility of this approach and provides insight to cAMP-dependent transcription. Conclusion: We demonstrate functional consequences of depleting multiple endogenous target genes using miR-shRNAs, and highlight the versatility of the described vector platform for multiple target gene knockdown in mammalian cells.

引用

页数：11

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