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Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference

RNA interference (RNAi) is a potent mechanism that silences mRNA and protein expression in all cells and tissue types. RNAi is known to exert many of its functional effects in the cytoplasm, and thus, the cellular localization of target mRNA may impact observed potency. Here, we demonstrate that cel...

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Autores principales: Ferguson, Chantal M., Echeverria, Dimas, Hassler, Matthew, Ly, Socheata, Khvorova, Anastasia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340969/
https://www.ncbi.nlm.nih.gov/pubmed/32650236
http://dx.doi.org/10.1016/j.omtn.2020.06.006
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author Ferguson, Chantal M.
Echeverria, Dimas
Hassler, Matthew
Ly, Socheata
Khvorova, Anastasia
author_facet Ferguson, Chantal M.
Echeverria, Dimas
Hassler, Matthew
Ly, Socheata
Khvorova, Anastasia
author_sort Ferguson, Chantal M.
collection PubMed
description RNA interference (RNAi) is a potent mechanism that silences mRNA and protein expression in all cells and tissue types. RNAi is known to exert many of its functional effects in the cytoplasm, and thus, the cellular localization of target mRNA may impact observed potency. Here, we demonstrate that cell identity has a profound impact on accessibility of apolipoprotein E (ApoE) mRNA to RNAi. We show that, whereas both neuronal and glial cell lines express detectable ApoE mRNA, in neuronal cells, ApoE mRNA is not targetable by RNAi. Screening of a panel of thirty-five chemically modified small interfering RNAs (siRNAs) did not produce a single hit in a neuronal cell line, whereas up to fifteen compounds showed strong efficacy in glial cells. Further investigation of the cellular localization of ApoE mRNA demonstrates that ApoE mRNA is partially spliced and preferentially localized to the nucleus (∼80%) in neuronal cells, whereas more than 90% of ApoE mRNA is cytoplasmic in glial cells. Such an inconsistency in intracellular localization and splicing might provide an explanation for functional differences in RNAi compounds. Thus, cellular origin might have an impact on accessibility of mRNA to RNAi and should be taken into account during the screening process.
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spelling pubmed-73409692020-07-14 Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference Ferguson, Chantal M. Echeverria, Dimas Hassler, Matthew Ly, Socheata Khvorova, Anastasia Mol Ther Nucleic Acids Article RNA interference (RNAi) is a potent mechanism that silences mRNA and protein expression in all cells and tissue types. RNAi is known to exert many of its functional effects in the cytoplasm, and thus, the cellular localization of target mRNA may impact observed potency. Here, we demonstrate that cell identity has a profound impact on accessibility of apolipoprotein E (ApoE) mRNA to RNAi. We show that, whereas both neuronal and glial cell lines express detectable ApoE mRNA, in neuronal cells, ApoE mRNA is not targetable by RNAi. Screening of a panel of thirty-five chemically modified small interfering RNAs (siRNAs) did not produce a single hit in a neuronal cell line, whereas up to fifteen compounds showed strong efficacy in glial cells. Further investigation of the cellular localization of ApoE mRNA demonstrates that ApoE mRNA is partially spliced and preferentially localized to the nucleus (∼80%) in neuronal cells, whereas more than 90% of ApoE mRNA is cytoplasmic in glial cells. Such an inconsistency in intracellular localization and splicing might provide an explanation for functional differences in RNAi compounds. Thus, cellular origin might have an impact on accessibility of mRNA to RNAi and should be taken into account during the screening process. American Society of Gene & Cell Therapy 2020-06-12 /pmc/articles/PMC7340969/ /pubmed/32650236 http://dx.doi.org/10.1016/j.omtn.2020.06.006 Text en © 2020 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ferguson, Chantal M.
Echeverria, Dimas
Hassler, Matthew
Ly, Socheata
Khvorova, Anastasia
Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title_full Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title_fullStr Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title_full_unstemmed Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title_short Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference
title_sort cell type impacts accessibility of mrna to silencing by rna interference
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7340969/
https://www.ncbi.nlm.nih.gov/pubmed/32650236
http://dx.doi.org/10.1016/j.omtn.2020.06.006
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