A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters
Name
nihms929464.pdf
Description
Accepted version
Size
2.27 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Piatkevich, Kiryl D
Jung, Erica E
Straub, Christoph
Linghu, Changyang
Park, Demian
Suk, Ho-Jun
Hochbaum, Daniel R
Goodwin, Daniel
Pnevmatikakis, Eftychios
Pak, Nikita
Date Issued
2018
Journal
Nature Chemical Biology
Publisher
Springer Science and Business Media LLC
Version
Author's final manuscript
Abstract
© 2018 The Author(s). We developed a new way to engineer complex proteins toward multidimensional specifications using a simple, yet scalable, directed evolution strategy. By robotically picking mammalian cells that were identified, under a microscope, as expressing proteins that simultaneously exhibit several specific properties, we can screen hundreds of thousands of proteins in a library in just a few hours, evaluating each along multiple performance axes. To demonstrate the power of this approach, we created a genetically encoded fluorescent voltage indicator, simultaneously optimizing its brightness and membrane localization using our microscopy-guided cell-picking strategy. We produced the high-performance opsin-based fluorescent voltage reporter Archon1 and demonstrated its utility by imaging spiking and millivolt-scale subthreshold and synaptic activity in acute mouse brain slices and in larval zebrafish in vivo. We also measured postsynaptic responses downstream of optogenetically controlled neurons in C. elegans.
MIT Department
Massachusetts Institute of Technology. Media Laboratory
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Mechanical Engineering
Picower Institute for Learning and Memory
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Massachusetts Institute of Technology. Department of Biological Engineering
McGovern Institute for Brain Research at MIT
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
Persistent DSpace Link
DOI of Published Version
10.1038/S41589-018-0004-9
https://doi.org/10.1038/S41589-018-0004-9
