PMID- 28045247
OWN - NLM
STAT- MEDLINE
DCOM- 20171025
LR  - 20190208
IS  - 1948-7193 (Electronic)
IS  - 1948-7193 (Linking)
VI  - 8
IP  - 3
DP  - 2017 Mar 15
TI  - Directed Evolution of Key Residues in Fluorescent Protein Inverses the Polarity 
      of Voltage Sensitivity in the Genetically Encoded Indicator ArcLight.
PG  - 513-523
LID - 10.1021/acschemneuro.6b00234 [doi]
AB  - Genetically encoded calcium indicators (GECIs) produce unprecedentedly large 
      signals that have enabled routine optical recording of single neuron activity in 
      vivo in rodent brain. Genetically encoded voltage indicators (GEVIs) offer a more 
      direct measure of neuronal electrical status, however the signal-to-noise 
      characteristics and signal polarity of the probes developed to date have 
      precluded routine use in vivo. We applied directed evolution to target modulable 
      areas of the fluorescent protein in GEVI ArcLight to create the first GFP-based 
      GEVI (Marina) that exhibits a DeltaF/DeltaV with a positive slope relationship. We found 
      that only three rounds of site-directed mutagenesis produced a family of 
      "brightening" GEVIs with voltage sensitivities comparable to that seen in the 
      parent probe ArcLight. This shift in signal polarity is an essential first step 
      to producing voltage indicators with signal-to-noise characteristics comparable 
      to GECIs to support widespread use in vivo.
FAU - Platisa, Jelena
AU  - Platisa J
AUID- ORCID: 0000-0003-4813-609X
AD  - The John B. Pierce Laboratory, Inc. , New Haven, Connecticut 06519, United 
      States.
FAU - Vasan, Ganesh
AU  - Vasan G
AD  - The John B. Pierce Laboratory, Inc. , New Haven, Connecticut 06519, United 
      States.
FAU - Yang, Amy
AU  - Yang A
AD  - The John B. Pierce Laboratory, Inc. , New Haven, Connecticut 06519, United 
      States.
FAU - Pieribone, Vincent A
AU  - Pieribone VA
AD  - The John B. Pierce Laboratory, Inc. , New Haven, Connecticut 06519, United 
      States.
LA  - eng
GR  - R01 NS083875/NS/NINDS NIH HHS/United States
GR  - U01 NS090565/NS/NINDS NIH HHS/United States
GR  - U01 NS103517/NS/NINDS NIH HHS/United States
GR  - UL1 TR001863/TR/NCATS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20170202
PL  - United States
TA  - ACS Chem Neurosci
JT  - ACS chemical neuroscience
JID - 101525337
RN  - 0 (Arclight fusion protein)
RN  - 0 (Luminescent Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Cerebral Cortex/cytology
MH  - Electric Stimulation
MH  - Embryo, Mammalian
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Luminescent Proteins/*chemistry/genetics/*metabolism
MH  - Mice
MH  - Models, Molecular
MH  - Molecular Biology
MH  - Mutagenesis, Site-Directed
MH  - Mutation/*genetics
MH  - Neurons/drug effects/*physiology
MH  - Patch-Clamp Techniques
MH  - Recombinant Fusion Proteins/*chemistry/genetics/*metabolism
MH  - Transfection
MH  - Voltage-Sensitive Dye Imaging
PMC - PMC5355904
OTO - NOTNLM
OT  - ArcLight
OT  - Genetically encoded voltage indicators
OT  - biosensors
OT  - directed evolution
OT  - green fluorescent protein
OT  - mutagenesis
COIS- The authors declare no competing financial interest.
EDAT- 2017/01/04 06:00
MHDA- 2017/10/27 06:00
PMCR- 2017/03/17
CRDT- 2017/01/04 06:00
PHST- 2017/01/04 06:00 [pubmed]
PHST- 2017/10/27 06:00 [medline]
PHST- 2017/01/04 06:00 [entrez]
PHST- 2017/03/17 00:00 [pmc-release]
AID - 10.1021/acschemneuro.6b00234 [doi]
PST - ppublish
SO  - ACS Chem Neurosci. 2017 Mar 15;8(3):513-523. doi: 10.1021/acschemneuro.6b00234. 
      Epub 2017 Feb 2.