| Enhanced target gene knockdown by a bifunctional shRNA: a novel approach of RNA interference. | |
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MedLine Citation:
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PMID: 20596090 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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RNA interference (RNAi) is a natural cellular regulatory process that inhibits gene expression by transcriptional, post-transcriptional and translational mechanisms. Synthetic approaches that emulate this process (small interfering RNA (siRNA), short hairpin RNA (shRNA)) have been shown to be similarly effective in this regard. We developed a novel 'bifunctional' RNAi strategy, which further optimizes target gene knockdown outcome. A bifunctional construct (bi-sh-STMN1) was generated against Stathmin1, a critical tubulin modulator that is overexpressed in human cancers. The bifunctional construct is postulated to concurrently repress the translation of the target mRNA (cleavage-independent, mRNA sequestration and degradation) and degrade (through RNase H-like cleavage) post-transcriptional mRNA through cleavage-dependent activities. Bi-sh-STMN1 showed enhanced potency and durability in parallel comparisons with conventional shRNA and siRNAs targeting the same sequence. Enhanced STMN1 protein knockdown by bi-sh-STMN1 was accompanied by target site cleavage at the mRNA level showed by the rapid amplification of complementary DNA ends (RACE) assay. Bi-sh-STMN1 also showed knockdown kinetics at the mRNA level consistent with its multieffector silencing mechanisms. The bifunctional shRNA is a highly effective and advantageous approach mediating RNAi at concentrations significantly lower than conventional shRNA or siRNA. These results support further evaluations. |
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Authors:
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D D Rao; P B Maples; N Senzer; P Kumar; Z Wang; B O Pappen; Y Yu; C Haddock; C Jay; A P Phadke; S Chen; J Kuhn; D Dylewski; S Scott; D Monsma; C Webb; A Tong; D Shanahan; J Nemunaitis |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-07-02 |
Journal Detail:
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Title: Cancer gene therapy Volume: 17 ISSN: 1476-5500 ISO Abbreviation: Cancer Gene Ther. Publication Date: 2010 Nov |
Date Detail:
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Created Date: 2010-10-18 Completed Date: 2011-01-27 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9432230 Medline TA: Cancer Gene Ther Country: England |
Other Details:
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Languages: eng Pagination: 780-91 Citation Subset: IM |
Affiliation:
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Gradalis, Inc., Dallas, TX, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Base Sequence Cell Line, Tumor Gene Knockdown Techniques / methods* Genetic Vectors Humans Molecular Sequence Data RNA Interference* RNA, Messenger / metabolism RNA, Small Interfering / metabolism* Stathmin / genetics, metabolism* Transcription, Genetic |
| Chemical | |
Reg. No./Substance:
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0/RNA, Messenger; 0/RNA, Small Interfering; 0/STMN1 protein, human; 0/Stathmin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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