PMID- 34326458
OWN - NLM
STAT- MEDLINE
DCOM- 20211111
LR  - 20240403
IS  - 2399-3642 (Electronic)
IS  - 2399-3642 (Linking)
VI  - 4
IP  - 1
DP  - 2021 Jul 29
TI  - Rational engineering of ratiometric calcium sensors with bright green and red 
      fluorescent proteins.
PG  - 924
LID - 10.1038/s42003-021-02452-z [doi]
LID - 924
AB  - Ratiometric genetically encoded calcium indicators (GECIs) record neural activity 
      with high brightness while mitigating motion-induced artifacts. Recently 
      developed ratiometric GECIs primarily employ cyan and yellow-fluorescent 
      fluorescence resonance energy transfer pairs, and thus fall short in some 
      applications that require deep tissue penetration and resistance to 
      photobleaching. We engineered a set of green-red ratiometric calcium sensors that 
      fused two fluorescent proteins and calcium sensing domain within an alternate 
      configuration. The best performing elements of this palette of sensors, Twitch-GR 
      and Twitch-NR, inherited the superior photophysical properties of their 
      constituent fluorescent proteins. These properties enabled our sensors to 
      outperform existing ratiometric calcium sensors in brightness and photobleaching 
      metrics. In turn, the shot-noise limited signal fidelity of our sensors when 
      reporting action potentials in cultured neurons and in the awake behaving mice 
      was higher than the fidelity of existing sensors. Our sensor enabled a regime of 
      imaging that simultaneously captured neural structure and function down to the 
      deep layers of the mouse cortex.
CI  - (c) 2021. The Author(s).
FAU - Zhang, Diming
AU  - Zhang D
AUID- ORCID: 0000-0001-6256-5638
AD  - Department of Biomedical Engineering, Duke University, Durham, NC, USA. 
      diming.zhang@duke.edu.
FAU - Redington, Emily
AU  - Redington E
AD  - Department of Biomedical Engineering, Duke University, Durham, NC, USA.
FAU - Gong, Yiyang
AU  - Gong Y
AUID- ORCID: 0000-0002-1855-930X
AD  - Department of Biomedical Engineering, Duke University, Durham, NC, USA. 
      yiyang.gong@duke.edu.
LA  - eng
SI  - figshare/10.6084/m9.figshare.14842467
GR  - DP2 NS111505/NS/NINDS NIH HHS/United States
GR  - UF1 NS107678/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20210729
PL  - England
TA  - Commun Biol
JT  - Communications biology
JID - 101719179
RN  - 0 (Intracellular Calcium-Sensing Proteins)
RN  - 0 (Luminescent Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Calcium/*chemistry
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Green Fluorescent Proteins/*chemistry
MH  - Intracellular Calcium-Sensing Proteins/*chemistry
MH  - Luminescent Proteins/*chemistry
MH  - *Protein Engineering
MH  - Red Fluorescent Protein
PMC - PMC8322158
COIS- The authors declare no competing interests.
EDAT- 2021/07/31 06:00
MHDA- 2021/11/12 06:00
PMCR- 2021/07/29
CRDT- 2021/07/30 06:32
PHST- 2021/01/26 00:00 [received]
PHST- 2021/07/14 00:00 [accepted]
PHST- 2021/07/30 06:32 [entrez]
PHST- 2021/07/31 06:00 [pubmed]
PHST- 2021/11/12 06:00 [medline]
PHST- 2021/07/29 00:00 [pmc-release]
AID - 10.1038/s42003-021-02452-z [pii]
AID - 2452 [pii]
AID - 10.1038/s42003-021-02452-z [doi]
PST - epublish
SO  - Commun Biol. 2021 Jul 29;4(1):924. doi: 10.1038/s42003-021-02452-z.