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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-09-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253120301670 |
| _version_ | 1819082421725822976 |
|---|---|
| author | Chantal M. Ferguson Dimas Echeverria Matthew Hassler Socheata Ly Anastasia Khvorova |
| author_facet | Chantal M. Ferguson Dimas Echeverria Matthew Hassler Socheata Ly Anastasia Khvorova |
| author_sort | Chantal M. Ferguson |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-21T20:16:25Z |
| format | Article |
| id | doaj.art-6a92b1c08c20454aad65ef9c7254dc4f |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-21T20:16:25Z |
| publishDate | 2020-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-6a92b1c08c20454aad65ef9c7254dc4f2022-12-21T18:51:36ZengElsevierMolecular Therapy: Nucleic Acids2162-25312020-09-0121384393Cell Type Impacts Accessibility of mRNA to Silencing by RNA InterferenceChantal M. Ferguson0Dimas Echeverria1Matthew Hassler2Socheata Ly3Anastasia Khvorova4RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USARNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USARNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USARNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USARNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA; Corresponding author: Anastasia Khvorova, University of Massachusetts Medical School, 368 Plantation Street, Worcester, MA 01605, USA.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.http://www.sciencedirect.com/science/article/pii/S2162253120301670 |
| spellingShingle | Chantal M. Ferguson Dimas Echeverria Matthew Hassler Socheata Ly Anastasia Khvorova Cell Type Impacts Accessibility of mRNA to Silencing by RNA Interference Molecular Therapy: Nucleic Acids |
| 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 |
| url | http://www.sciencedirect.com/science/article/pii/S2162253120301670 |
| work_keys_str_mv | AT chantalmferguson celltypeimpactsaccessibilityofmrnatosilencingbyrnainterference AT dimasecheverria celltypeimpactsaccessibilityofmrnatosilencingbyrnainterference AT matthewhassler celltypeimpactsaccessibilityofmrnatosilencingbyrnainterference AT socheataly celltypeimpactsaccessibilityofmrnatosilencingbyrnainterference AT anastasiakhvorova celltypeimpactsaccessibilityofmrnatosilencingbyrnainterference |