Show simple item record

dc.contributor.authorHasegawa, Yuto
dc.contributor.authorMao, Wenjie
dc.contributor.authorSaha, Sucharita
dc.contributor.authorGunner, Georgia
dc.contributor.authorKolpakova, Jenya
dc.contributor.authorMartin, Gilles E.
dc.contributor.authorFutai, Kensuke
dc.date2022-08-11T08:08:22.000
dc.date.accessioned2022-08-23T15:52:48Z
dc.date.available2022-08-23T15:52:48Z
dc.date.issued2017-10-11
dc.date.submitted2018-01-22
dc.identifier.citation<p>eNeuro. 2017 Oct 11;4(5). pii: ENEURO.0186-17.2017. doi: 10.1523/ENEURO.0186-17.2017. eCollection 2017 Sep-Oct. <a href="https://doi.org/10.1523/ENEURO.0186-17.2017">Link to article on publisher's site</a></p>
dc.identifier.issn2373-2822 (Linking)
dc.identifier.doi10.1523/ENEURO.0186-17.2017
dc.identifier.pmid29034317
dc.identifier.urihttp://hdl.handle.net/20.500.14038/29221
dc.description.abstractRNA interference (RNAi) is a straightforward approach to study gene function from the in vitro cellular level to in vivo animal behavior. Although RNAi-mediated gene knockdown has become essentially routine in neuroscience over the past ten years, off-target effects of short hairpin RNAs (shRNAs) should be considered as the proper choice of control shRNA is critical in order to perform meaningful experiments. Luciferase shRNA (shLuc), targeting firefly luciferase, and scrambled shRNAs (shScrs) have been widely used as controls for vertebrate cell research. However, thorough validation of control shRNAs has not been made to date. Here, we performed thorough physiological and morphological studies against control shRNAs in mouse hippocampal CA1 pyramidal neurons. As expected, all control shRNAs exhibited normal basal synaptic transmission and dendritic morphology. However, to our surprise, shLuc exerted severe off-target effects on voltage-gated ion channel function, while the shScr had no detectable changes. These results indicate that thorough validation of shRNA is imperative and, in the absence of such validation, that shScr is the best available negative control for gene knockdown studies.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=29034317&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rightsCopyright © 2017 Hasegawa et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjection channels
dc.subjectluciferase
dc.subjectoff-target effects
dc.subjectRNA interference
dc.subjectshort hairpin
dc.subjectNeuroscience and Neurobiology
dc.titleLuciferase shRNA Presents off-Target Effects on Voltage-Gated Ion Channels in Mouse Hippocampal Pyramidal Neurons
dc.typeJournal Article
dc.source.journaltitleeNeuro
dc.source.volume4
dc.source.issue5
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2456&amp;context=faculty_pubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/faculty_pubs/1453
dc.identifier.contextkey11398484
refterms.dateFOA2022-08-23T15:52:48Z
html.description.abstract<p>RNA interference (RNAi) is a straightforward approach to study gene function from the in vitro cellular level to in vivo animal behavior. Although RNAi-mediated gene knockdown has become essentially routine in neuroscience over the past ten years, off-target effects of short hairpin RNAs (shRNAs) should be considered as the proper choice of control shRNA is critical in order to perform meaningful experiments. Luciferase shRNA (shLuc), targeting firefly luciferase, and scrambled shRNAs (shScrs) have been widely used as controls for vertebrate cell research. However, thorough validation of control shRNAs has not been made to date. Here, we performed thorough physiological and morphological studies against control shRNAs in mouse hippocampal CA1 pyramidal neurons. As expected, all control shRNAs exhibited normal basal synaptic transmission and dendritic morphology. However, to our surprise, shLuc exerted severe off-target effects on voltage-gated ion channel function, while the shScr had no detectable changes. These results indicate that thorough validation of shRNA is imperative and, in the absence of such validation, that shScr is the best available negative control for gene knockdown studies.</p>
dc.identifier.submissionpathfaculty_pubs/1453
dc.contributor.departmentNeurobiology
dc.contributor.departmentGraduate School of Biomedical Sciences, Neuroscience Program
dc.contributor.departmentFutai Lab
dc.contributor.departmentMartin Lab
dc.contributor.departmentBrudnick Neuropsychiatric Research Institute
dc.contributor.studentWenjie Mao
dc.contributor.studentJenya Kolpakova
dc.contributor.studentGeorgia Gunner
dc.description.thesisprogramNeuroscience


Files in this item

Thumbnail
Name:
ENEURO.0186_17.2017.pdf
Size:
2.012Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record

Copyright © 2017 Hasegawa et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Except where otherwise noted, this item's license is described as Copyright © 2017 Hasegawa et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.