Luciferase shRNA Presents off-Target Effects on Voltage-Gated Ion Channels in Mouse Hippocampal Pyramidal Neurons
Martin, Gilles E.
UMass Chan AffiliationsNeurobiology
Graduate School of Biomedical Sciences, Neuroscience Program
Brudnick Neuropsychiatric Research Institute
Document TypeJournal Article
Neuroscience and Neurobiology
MetadataShow full item record
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.
eNeuro. 2017 Oct 11;4(5). pii: ENEURO.0186-17.2017. doi: 10.1523/ENEURO.0186-17.2017. eCollection 2017 Sep-Oct. Link to article on publisher's site
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/29221
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.
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.