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RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA

Twenty one base pair long small interfering RNAs (siRNAs) are widely in use in mammalian RNAi experiments. The present study assesses the capability of 43 and 63bp dsRNAs with two 2nt long 3′-overhangs to induce RNAi in mammalian and Drosophila cells. Human Dicer was found to cleave these dsRNAs fro...

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Autores principales: Ui-Tei, Kumiko, Zenno, Shuhei, Juni, Aya, Saigo, Kaoru
Formato: Texto
Lenguaje:English
Publicado: Library Publishing Media 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737202/
https://www.ncbi.nlm.nih.gov/pubmed/19771208
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author Ui-Tei, Kumiko
Zenno, Shuhei
Juni, Aya
Saigo, Kaoru
author_facet Ui-Tei, Kumiko
Zenno, Shuhei
Juni, Aya
Saigo, Kaoru
author_sort Ui-Tei, Kumiko
collection PubMed
description Twenty one base pair long small interfering RNAs (siRNAs) are widely in use in mammalian RNAi experiments. The present study assesses the capability of 43 and 63bp dsRNAs with two 2nt long 3′-overhangs to induce RNAi in mammalian and Drosophila cells. Human Dicer was found to cleave these dsRNAs from their ends to generate two or three monomeric siRNA units, each 21-22bp in length. When, in 43bp dsRNA, there was present a highly-effective siRNA sequence in frame with respect to the Dicer digestion, considerably high RNAi activity was noted to be induced in mouse embryonic stem E14TG2a, human HeLa, Chinese hamster CHO-K1 or Drosophila S2 cells. In contrast, RNAi depending on 63bp dsRNA, containing a highly effective siRNA sequence in frame with respect to Dicer digestion, varied considerably depending on cell lines used. While there was no appreciable RNAi in HeLa cells associated with relatively strong interferon response, a significant level of RNAi was noted in E14TG2a, CHO-K1 and S2 cells, in all of which interferon response induction was but slight, if at all. It would thus follow that siRNA oligomers with sequence of a highly functional siRNA monomer unit in frame with respect to dicer digestion should serve as a good RNAi agent in Drosophila and certain mammalian cells.
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spelling pubmed-27372022009-09-21 RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA Ui-Tei, Kumiko Zenno, Shuhei Juni, Aya Saigo, Kaoru J RNAi Gene Silencing Research Article Twenty one base pair long small interfering RNAs (siRNAs) are widely in use in mammalian RNAi experiments. The present study assesses the capability of 43 and 63bp dsRNAs with two 2nt long 3′-overhangs to induce RNAi in mammalian and Drosophila cells. Human Dicer was found to cleave these dsRNAs from their ends to generate two or three monomeric siRNA units, each 21-22bp in length. When, in 43bp dsRNA, there was present a highly-effective siRNA sequence in frame with respect to the Dicer digestion, considerably high RNAi activity was noted to be induced in mouse embryonic stem E14TG2a, human HeLa, Chinese hamster CHO-K1 or Drosophila S2 cells. In contrast, RNAi depending on 63bp dsRNA, containing a highly effective siRNA sequence in frame with respect to Dicer digestion, varied considerably depending on cell lines used. While there was no appreciable RNAi in HeLa cells associated with relatively strong interferon response, a significant level of RNAi was noted in E14TG2a, CHO-K1 and S2 cells, in all of which interferon response induction was but slight, if at all. It would thus follow that siRNA oligomers with sequence of a highly functional siRNA monomer unit in frame with respect to dicer digestion should serve as a good RNAi agent in Drosophila and certain mammalian cells. Library Publishing Media 2005-10-14 /pmc/articles/PMC2737202/ /pubmed/19771208 Text en © Copyright Kumiko Ui-Tei et al http://www.libpubmedia.co.uk/RNAiJ/LicenceForUsers.pdf This is an open access article, published under the terms of the Licence for Users available at http://www.libpubmedia.co.uk/RNAiJ/LicenceForUsers.pdf. This licence permits non-commercial use, distribution and reproduction of the article, provided the original work is appropriately acknowledged with correct citation details.
spellingShingle Research Article
Ui-Tei, Kumiko
Zenno, Shuhei
Juni, Aya
Saigo, Kaoru
RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title_full RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title_fullStr RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title_full_unstemmed RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title_short RNAi induced in mammalian and Drosophila cells via transfection of dimers and trimers of small interfering RNA
title_sort rnai induced in mammalian and drosophila cells via transfection of dimers and trimers of small interfering rna
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737202/
https://www.ncbi.nlm.nih.gov/pubmed/19771208
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