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A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters
Authors
A Currin
A Gordus
+85 more
A Subedi
AM Stewart
AS Chuong
Balint Szabo
Bernardo L. Sabatini
BL Fiedler
BY Chow
BY Chow
C Chen
C Wyart
Changyang Linghu
Chao-Tsung Yang
Christoph Straub
Clemens Riegler
Daniel Goodwin
Daniel R. Hochbaum
DD Lee
Demian Park
DR Hochbaum
E Giraud
Edward S. Boyden
Eftychios Pnevmatikakis
Erica E. Jung
ES Boyden
F St-Pierre
Florian Engert
FV Subach
G Grigoryan
G Pucihar
GS Filonov
H-W Ai
Ho-Jun Suk
JD Knowles
Jeffrey L. Rhoades
Jessica L. Saulnier
JM Kralj
JS Lebkowski
JY Qin
KD Piatkevich
Kiryl D. Piatkevich
KM Dean
KM Kwan
L He
Limor Freifeld
M Drobizhev
M Jiang
M Krzywinski
M Okazaki
MB Ahrens
MB Ahrens
MB Bouchard
Mikhail Drobizhev
Misha B. Ahrens
MJ Mahon
MS Packer
MZ Lin
NC Dobes
NC Flytzanis
NC Klapoetke
Nikita Pak
NS Makarov
O Renaud
OA Shemesh
Or Shemesh
R Prevedel
R Salánki
RB Dell
Re,AM Del
RS McIsaac
RW Friedrich
S Brenner
S Fisher
Shoh Asano
Steven W. Flavell
T Kotnik
T-W Chen
Takashi Kawashima
Thom Hughes
V Gradinaru
Y Gong
Y Gong
Y Kimura
Y Zhao
Young-Gyu Yoon
Z Környei
Publication date
19 July 2019
Publisher
'Springer Science and Business Media LLC'
Doi
Cite
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
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oai:dspace.mit.edu:1721.1/1346...
Last time updated on 19/12/2021
Crossref
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info:doi/10.1038%2Fs41589-018-...
Last time updated on 08/12/2020