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Endogenous Piezo1 Can Confound Mechanically Activated Channel Identification and Characterization

Abstract : A gold standard for characterizing mechanically activated (MA) currents is via heterologous expression of candidate channels in naive cells. Two recent studies described MA channels using this paradigm. TMEM150c was proposed to be a component of an MA channel partly based on a heterologous expression approach (Hong et al., 2016). In another study, Piezo1's N-terminal ``propeller'' domain was proposed to constitute an intrinsic mechanosensitive module based on expression of a chimera between a pore-forming domain of the mechanically insensitive ASIC1 channel and Piezo1 (Zhao et al., 2016). When we attempted to replicate these results, we found each construct conferred modest MA currents in a small fraction of naive HEK cells similar to the published work. Strikingly, these MA currents were not detected in cells in which endogenous Piezo1 was CRISPR/Cas9 inactivated. These results highlight the importance of choosing cells lacking endogenous MA channels to assay the mechanotransduction properties of various proteins. This Matters Arising paper is in response to Hong et al. (2016) and Zhao et al. (2016) in Neuron. See also the response papers by Hong et al. (2017) and Zhao et al. (2017) published concurrently with this Matters Arising.
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Submitted on : Friday, October 26, 2018 - 10:50:03 AM
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Adrienne E. Dubin, Swetha Murthy, Amanda H. Lewis, Lucie Brosse, Stuart M. Cahalan, et al.. Endogenous Piezo1 Can Confound Mechanically Activated Channel Identification and Characterization. Neuron, Elsevier, 2017, 94 (2), pp.266+. ⟨10.1016/j.neuron.2017.03.039⟩. ⟨hal-01765080⟩



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