A robotic multidimensional directed evolution approach applied to fluorescent voltage reporters

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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.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2018-04
Language
English
Article Type
Article
Citation

NATURE CHEMICAL BIOLOGY, v.14, no.4, pp.352 - +

ISSN
1552-4450
DOI
10.1038/s41589-018-0004-9
URI
http://hdl.handle.net/10203/246956
Appears in Collection
EE-Journal Papers(저널논문)
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