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Approaches for chemically synthesized siRNA and vector-mediated RNAi
Successful applications of RNAi in mammalian cells depend upon effective knockdown of targeted transcripts and efficient intracellular delivery of either preformed si/shRNAs or vector expressed si/shRNAs. We have previously demonstrated that 27 base pair double stranded RNAs which are substrates for...
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Published in: | FEBS letters 2005-10, Vol.579 (26), p.5974-5981 |
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creator | Amarzguioui, Mohammed Rossi, John J. Kim, Dongho |
description | Successful applications of RNAi in mammalian cells depend upon effective knockdown of targeted transcripts and efficient intracellular delivery of either preformed si/shRNAs or vector expressed si/shRNAs. We have previously demonstrated that 27 base pair double stranded RNAs which are substrates for Dicer can be up to 100 times more potent than 21mer siRNAs. In this mini-review we elaborate upon the rationale and design strategies for creating Dicer substrate RNAs that provide enhanced knockdown of targeted RNAs and minimize the utilization of the sense strand as RNAi effectors. Expression of shRNAs or siRNAs in mammalian cells can be achieved via transcription from either Pol II or Pol III promoters. There are certain constrictions in designing such vectors, and these are described here. Additionally, we review strategies for inducible shRNA expression and the various viral vectors that can be used to transduce shRNA genes into a variety of cells and tissues. The overall goal of this mini-review is to provide an overview of available approaches for optimizing RNAi mediated down regulation of gene expression in mammalian cells via RNA interference. Although the primary focus is the use of RNAi mediated cleavage of targeted transcripts, it is highly probable that some of the approaches described herein will be applicable to RNAi mediated inhibition of translation and transcriptional gene silencing. |
doi_str_mv | 10.1016/j.febslet.2005.08.070 |
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We have previously demonstrated that 27 base pair double stranded RNAs which are substrates for Dicer can be up to 100 times more potent than 21mer siRNAs. In this mini-review we elaborate upon the rationale and design strategies for creating Dicer substrate RNAs that provide enhanced knockdown of targeted RNAs and minimize the utilization of the sense strand as RNAi effectors. Expression of shRNAs or siRNAs in mammalian cells can be achieved via transcription from either Pol II or Pol III promoters. There are certain constrictions in designing such vectors, and these are described here. Additionally, we review strategies for inducible shRNA expression and the various viral vectors that can be used to transduce shRNA genes into a variety of cells and tissues. The overall goal of this mini-review is to provide an overview of available approaches for optimizing RNAi mediated down regulation of gene expression in mammalian cells via RNA interference. 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We have previously demonstrated that 27 base pair double stranded RNAs which are substrates for Dicer can be up to 100 times more potent than 21mer siRNAs. In this mini-review we elaborate upon the rationale and design strategies for creating Dicer substrate RNAs that provide enhanced knockdown of targeted RNAs and minimize the utilization of the sense strand as RNAi effectors. Expression of shRNAs or siRNAs in mammalian cells can be achieved via transcription from either Pol II or Pol III promoters. There are certain constrictions in designing such vectors, and these are described here. Additionally, we review strategies for inducible shRNA expression and the various viral vectors that can be used to transduce shRNA genes into a variety of cells and tissues. The overall goal of this mini-review is to provide an overview of available approaches for optimizing RNAi mediated down regulation of gene expression in mammalian cells via RNA interference. Although the primary focus is the use of RNAi mediated cleavage of targeted transcripts, it is highly probable that some of the approaches described herein will be applicable to RNAi mediated inhibition of translation and transcriptional gene silencing.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Dicer</subject><subject>Gene Silencing</subject><subject>Gene Transfer Techniques</subject><subject>Genetic Techniques</subject><subject>Genetic Vectors</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Humans</subject><subject>Inducible shRNA expression</subject><subject>MicroRNAs - chemistry</subject><subject>Models, Genetic</subject><subject>Molecular Sequence Data</subject><subject>Pol II</subject><subject>Pol III</subject><subject>Promoter Regions, Genetic</subject><subject>Protein Structure, Tertiary</subject><subject>PTGS</subject><subject>Ribonuclease III - genetics</subject><subject>RNA - metabolism</subject><subject>RNA Interference - physiology</subject><subject>RNA Polymerase III - chemistry</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Small Interfering - metabolism</subject><subject>siRNA</subject><subject>Transcription, Genetic</subject><subject>Viral vector</subject><issn>0014-5793</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><recordid>eNqNkV9PwjAUxRujEUQ_gmZPvm3edrTrngwiiAnRxD_PTdfdxZLBsB0Y_PQWIfERn9rennt6-zuEXFJIKFBxM0sqLHyNbcIAeAIygQyOSJfKLI3TvpDHpAtA-zHP8rRDzryfQThLmp-SDhU0zyGVXXI_WC5do80H-qhqXBQ2c2t0XW8iv1m0oWy_sYy8fXkaRHpRRms0bePiOZZWt-Em1O05Oal07fFiv_bI-3j0NpzE0-eHx-FgGhueMRYzBIlMlyKVBStoWgDmEoQBQJpVIDkTpehXhhfcAApDKwa51BmX4aul0GmPXO98w8ifK_StmltvsK71ApuVV0JmrJ9SelDIgIEAYEHId0LjGu8dVmrp7Fy7jaKgtpzVTO05qy1nBVIFzqHvav_Aqggo_rr2YINgshN82Ro3_3NV49Ede92Gts0MOFDIf2e83VlhQLu26JQ3FhcmJOBCGKps7IFpfwCHHqXD</recordid><startdate>20051031</startdate><enddate>20051031</enddate><creator>Amarzguioui, Mohammed</creator><creator>Rossi, John J.</creator><creator>Kim, Dongho</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20051031</creationdate><title>Approaches for chemically synthesized siRNA and vector-mediated RNAi</title><author>Amarzguioui, Mohammed ; Rossi, John J. ; Kim, Dongho</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5722-2e08e2ad638b2b13b0e9806c00e17f08526d64fc5b5c0e6c1f2098a758febd6a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Animals</topic><topic>Base Sequence</topic><topic>Dicer</topic><topic>Gene Silencing</topic><topic>Gene Transfer Techniques</topic><topic>Genetic Techniques</topic><topic>Genetic Vectors</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Humans</topic><topic>Inducible shRNA expression</topic><topic>MicroRNAs - chemistry</topic><topic>Models, Genetic</topic><topic>Molecular Sequence Data</topic><topic>Pol II</topic><topic>Pol III</topic><topic>Promoter Regions, Genetic</topic><topic>Protein Structure, Tertiary</topic><topic>PTGS</topic><topic>Ribonuclease III - genetics</topic><topic>RNA - metabolism</topic><topic>RNA Interference - physiology</topic><topic>RNA Polymerase III - chemistry</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Small Interfering - metabolism</topic><topic>siRNA</topic><topic>Transcription, Genetic</topic><topic>Viral vector</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Amarzguioui, Mohammed</creatorcontrib><creatorcontrib>Rossi, John J.</creatorcontrib><creatorcontrib>Kim, Dongho</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Amarzguioui, Mohammed</au><au>Rossi, John J.</au><au>Kim, Dongho</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Approaches for chemically synthesized siRNA and vector-mediated RNAi</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>2005-10-31</date><risdate>2005</risdate><volume>579</volume><issue>26</issue><spage>5974</spage><epage>5981</epage><pages>5974-5981</pages><issn>0014-5793</issn><eissn>1873-3468</eissn><abstract>Successful applications of RNAi in mammalian cells depend upon effective knockdown of targeted transcripts and efficient intracellular delivery of either preformed si/shRNAs or vector expressed si/shRNAs. We have previously demonstrated that 27 base pair double stranded RNAs which are substrates for Dicer can be up to 100 times more potent than 21mer siRNAs. In this mini-review we elaborate upon the rationale and design strategies for creating Dicer substrate RNAs that provide enhanced knockdown of targeted RNAs and minimize the utilization of the sense strand as RNAi effectors. Expression of shRNAs or siRNAs in mammalian cells can be achieved via transcription from either Pol II or Pol III promoters. There are certain constrictions in designing such vectors, and these are described here. Additionally, we review strategies for inducible shRNA expression and the various viral vectors that can be used to transduce shRNA genes into a variety of cells and tissues. The overall goal of this mini-review is to provide an overview of available approaches for optimizing RNAi mediated down regulation of gene expression in mammalian cells via RNA interference. 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subjects | Animals Base Sequence Dicer Gene Silencing Gene Transfer Techniques Genetic Techniques Genetic Vectors Green Fluorescent Proteins - metabolism Humans Inducible shRNA expression MicroRNAs - chemistry Models, Genetic Molecular Sequence Data Pol II Pol III Promoter Regions, Genetic Protein Structure, Tertiary PTGS Ribonuclease III - genetics RNA - metabolism RNA Interference - physiology RNA Polymerase III - chemistry RNA, Messenger - metabolism RNA, Small Interfering - metabolism siRNA Transcription, Genetic Viral vector |
title | Approaches for chemically synthesized siRNA and vector-mediated RNAi |
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