Cargando…

Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity

In both the pharmaceutical and agricultural fields, RNA-based products have capitalized upon the mechanism of RNA interference for targeted reduction of gene expression to improve phenotypes and traits. Reduction in gene expression by RNAi is the result of a small interfering RNA (siRNA) molecule bi...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Peizhen, Havecker, Ericka, Bauer, Matthew, Diehl, Carl, Hendrix, Bill, Hoffer, Paul, Boyle, Timothy, Bradley, John, Caruano-Yzermans, Amy, Deikman, Jill
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8423273/
https://www.ncbi.nlm.nih.gov/pubmed/34492058
http://dx.doi.org/10.1371/journal.pone.0256863
_version_ 1783749432692441088
author Yang, Peizhen
Havecker, Ericka
Bauer, Matthew
Diehl, Carl
Hendrix, Bill
Hoffer, Paul
Boyle, Timothy
Bradley, John
Caruano-Yzermans, Amy
Deikman, Jill
author_facet Yang, Peizhen
Havecker, Ericka
Bauer, Matthew
Diehl, Carl
Hendrix, Bill
Hoffer, Paul
Boyle, Timothy
Bradley, John
Caruano-Yzermans, Amy
Deikman, Jill
author_sort Yang, Peizhen
collection PubMed
description In both the pharmaceutical and agricultural fields, RNA-based products have capitalized upon the mechanism of RNA interference for targeted reduction of gene expression to improve phenotypes and traits. Reduction in gene expression by RNAi is the result of a small interfering RNA (siRNA) molecule binding to an ARGONAUTE (AGO) protein and directing the effector complex to a homologous region of a target gene’s mRNA. siRNAs properties that govern RNA-AGO association have been studied in detail. The siRNA 5’ nucleotide (nt) identity has been demonstrated in plants to be an important property responsible for directing association of endogenous small RNAs with different AGO effector proteins. However, it has not been investigated whether the 5’ nt identity is an efficacious determinant for topically-applied chemically synthesized siRNAs. In this study, we employed a sandpaper abrasion method to study the silencing efficacies of topically-applied 21 base-pair siRNA duplexes. The MAGNESIUM CHELATASE and GREEN FLUORESCENT PROTEIN genes were selected as endogenous and transgenic gene targets, respectively, to assess the molecular and phenotypic effects of gene silencing. Collections of siRNA variants with different 5’ nt identities and different pairing states between the 5’ antisense nt and its match in the sense strand of the siRNA duplex were tested for their silencing efficacy. Our results suggest a flexibility in the 5’ nt requirement for topically applied siRNA duplexes in planta and highlight the similarity of 5’ thermodynamic rules governing topical siRNA efficacy across plants and animals.
format Online
Article
Text
id pubmed-8423273
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-84232732021-09-08 Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity Yang, Peizhen Havecker, Ericka Bauer, Matthew Diehl, Carl Hendrix, Bill Hoffer, Paul Boyle, Timothy Bradley, John Caruano-Yzermans, Amy Deikman, Jill PLoS One Research Article In both the pharmaceutical and agricultural fields, RNA-based products have capitalized upon the mechanism of RNA interference for targeted reduction of gene expression to improve phenotypes and traits. Reduction in gene expression by RNAi is the result of a small interfering RNA (siRNA) molecule binding to an ARGONAUTE (AGO) protein and directing the effector complex to a homologous region of a target gene’s mRNA. siRNAs properties that govern RNA-AGO association have been studied in detail. The siRNA 5’ nucleotide (nt) identity has been demonstrated in plants to be an important property responsible for directing association of endogenous small RNAs with different AGO effector proteins. However, it has not been investigated whether the 5’ nt identity is an efficacious determinant for topically-applied chemically synthesized siRNAs. In this study, we employed a sandpaper abrasion method to study the silencing efficacies of topically-applied 21 base-pair siRNA duplexes. The MAGNESIUM CHELATASE and GREEN FLUORESCENT PROTEIN genes were selected as endogenous and transgenic gene targets, respectively, to assess the molecular and phenotypic effects of gene silencing. Collections of siRNA variants with different 5’ nt identities and different pairing states between the 5’ antisense nt and its match in the sense strand of the siRNA duplex were tested for their silencing efficacy. Our results suggest a flexibility in the 5’ nt requirement for topically applied siRNA duplexes in planta and highlight the similarity of 5’ thermodynamic rules governing topical siRNA efficacy across plants and animals. Public Library of Science 2021-09-07 /pmc/articles/PMC8423273/ /pubmed/34492058 http://dx.doi.org/10.1371/journal.pone.0256863 Text en © 2021 Yang et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Yang, Peizhen
Havecker, Ericka
Bauer, Matthew
Diehl, Carl
Hendrix, Bill
Hoffer, Paul
Boyle, Timothy
Bradley, John
Caruano-Yzermans, Amy
Deikman, Jill
Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title_full Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title_fullStr Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title_full_unstemmed Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title_short Beyond identity: Understanding the contribution of the 5’ nucleotide of the antisense strand to RNAi activity
title_sort beyond identity: understanding the contribution of the 5’ nucleotide of the antisense strand to rnai activity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8423273/
https://www.ncbi.nlm.nih.gov/pubmed/34492058
http://dx.doi.org/10.1371/journal.pone.0256863
work_keys_str_mv AT yangpeizhen beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT haveckerericka beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT bauermatthew beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT diehlcarl beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT hendrixbill beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT hofferpaul beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT boyletimothy beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT bradleyjohn beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT caruanoyzermansamy beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity
AT deikmanjill beyondidentityunderstandingthecontributionofthe5nucleotideoftheantisensestrandtornaiactivity