Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells

Abstract RNA interference (RNAi) is induced by the direct transfer of double-stranded RNAs (dsRNAs) into protoplasts prepared from Arabidopsis thaliana seedlings. In this protoplast RNAi system, we compared the efficacies of various-sized dsRNAs (between 21 and 139 nucleotides [nt]) for inducing RNA...

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Autores principales:	Sayaka Kakiyama, Midori Tabara, Yuki Nishibori, Hiromitsu Moriyama, Toshiyuki Fukuhara
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/866880b22db64a64acafaa91c69725b6
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spelling	oai:doaj.org-article:866880b22db64a64acafaa91c69725b62021-12-02T16:07:52ZLong DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells10.1038/s41598-019-43443-92045-2322https://doaj.org/article/866880b22db64a64acafaa91c69725b62019-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-43443-9https://doaj.org/toc/2045-2322Abstract RNA interference (RNAi) is induced by the direct transfer of double-stranded RNAs (dsRNAs) into protoplasts prepared from Arabidopsis thaliana seedlings. In this protoplast RNAi system, we compared the efficacies of various-sized dsRNAs (between 21 and 139 nucleotides [nt]) for inducing RNAi and assessed the dsRNA-cleaving activities of Dicer-like 3 (DCL3) and 4 (DCL4). After the direct transfer of dsRNAs into protoplasts, cleaved RNA products of 21 nt were detected from long 130- or 500-nt dsRNAs by DCL4 but not from 37-nt dsRNAs. These results indicate that DCL4 preferentially cleaves long dsRNAs in protoplasts, consistent with our previous biochemical data regarding the substrate specificity of DCL4. Direct transfer of long dsRNAs of approximately 130 nt into protoplasts induces RNAi much more effectively (by approximately 60- to 400-fold) than direct transfer of short 37-nt dsRNAs. Although transfer of 21-nt dsRNAs into protoplasts induced RNAi without DCL4 activity, the induction of RNAi was less effective (by approximately 0.01-fold) compared with long dsRNAs. These results indicate that cleavage of long dsRNAs exceeding 100 nt by DCL4 into 21-nt dsRNAs is essential for efficient induction of RNAi in plant cells.Sayaka KakiyamaMidori TabaraYuki NishiboriHiromitsu MoriyamaToshiyuki FukuharaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-10 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sayaka Kakiyama Midori Tabara Yuki Nishibori Hiromitsu Moriyama Toshiyuki Fukuhara Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
description	Abstract RNA interference (RNAi) is induced by the direct transfer of double-stranded RNAs (dsRNAs) into protoplasts prepared from Arabidopsis thaliana seedlings. In this protoplast RNAi system, we compared the efficacies of various-sized dsRNAs (between 21 and 139 nucleotides [nt]) for inducing RNAi and assessed the dsRNA-cleaving activities of Dicer-like 3 (DCL3) and 4 (DCL4). After the direct transfer of dsRNAs into protoplasts, cleaved RNA products of 21 nt were detected from long 130- or 500-nt dsRNAs by DCL4 but not from 37-nt dsRNAs. These results indicate that DCL4 preferentially cleaves long dsRNAs in protoplasts, consistent with our previous biochemical data regarding the substrate specificity of DCL4. Direct transfer of long dsRNAs of approximately 130 nt into protoplasts induces RNAi much more effectively (by approximately 60- to 400-fold) than direct transfer of short 37-nt dsRNAs. Although transfer of 21-nt dsRNAs into protoplasts induced RNAi without DCL4 activity, the induction of RNAi was less effective (by approximately 0.01-fold) compared with long dsRNAs. These results indicate that cleavage of long dsRNAs exceeding 100 nt by DCL4 into 21-nt dsRNAs is essential for efficient induction of RNAi in plant cells.
format	article
author	Sayaka Kakiyama Midori Tabara Yuki Nishibori Hiromitsu Moriyama Toshiyuki Fukuhara
author_facet	Sayaka Kakiyama Midori Tabara Yuki Nishibori Hiromitsu Moriyama Toshiyuki Fukuhara
author_sort	Sayaka Kakiyama
title	Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
title_short	Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
title_full	Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
title_fullStr	Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
title_full_unstemmed	Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells
title_sort	long dcl4-substrate dsrnas efficiently induce rna interference in plant cells
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/866880b22db64a64acafaa91c69725b6
work_keys_str_mv	AT sayakakakiyama longdcl4substratedsrnasefficientlyinducernainterferenceinplantcells AT midoritabara longdcl4substratedsrnasefficientlyinducernainterferenceinplantcells AT yukinishibori longdcl4substratedsrnasefficientlyinducernainterferenceinplantcells AT hiromitsumoriyama longdcl4substratedsrnasefficientlyinducernainterferenceinplantcells AT toshiyukifukuhara longdcl4substratedsrnasefficientlyinducernainterferenceinplantcells
_version_	1718384669020389376

Long DCL4-substrate dsRNAs efficiently induce RNA interference in plant cells

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