PMID- 26382605
OWN - NLM
STAT- MEDLINE
DCOM- 20160607
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 9
DP  - 2015
TI  - Green-to-Red Photoconversion of GCaMP.
PG  - e0138127
LID - 10.1371/journal.pone.0138127 [doi]
LID - e0138127
AB  - Genetically encoded calcium indicators (GECIs) permit imaging intracellular 
      calcium transients. Among GECIs, the GFP-based GCaMPs are the most widely used 
      because of their high sensitivity and rapid response to changes in intracellular 
      calcium concentrations. Here we report that the fluorescence of GCaMPs--including 
      GCaMP3, GCaMP5 and GCaMP6--can be converted from green to red following exposure 
      to blue-green light (450-500 nm). This photoconversion occurs in both insect and 
      mammalian cells and is enhanced in a low oxygen environment. The red fluorescent 
      GCaMPs retained calcium responsiveness, albeit with reduced sensitivity. We 
      identified several amino acid residues in GCaMP important for photoconversion and 
      generated a GCaMP variant with increased photoconversion efficiency in cell 
      culture. This light-induced spectral shift allows the ready labeling of specific, 
      targeted sets of GCaMP-expressing cells for functional imaging in the red 
      channel. Together, these findings indicate the potential for greater utility of 
      existing GCaMP reagents, including transgenic animals.
FAU - Ai, Minrong
AU  - Ai M
AD  - Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York 
      University, School of Medicine, New York, New York, United States of America.
FAU - Mills, Holly
AU  - Mills H
AD  - Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York 
      University, School of Medicine, New York, New York, United States of America.
FAU - Kanai, Makoto
AU  - Kanai M
AD  - Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York 
      University, School of Medicine, New York, New York, United States of America.
FAU - Lai, Jason
AU  - Lai J
AD  - Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York 
      University, School of Medicine, New York, New York, United States of America.
FAU - Deng, Jingjing
AU  - Deng J
AD  - Department of Biochemistry and Molecular Pharmacology, Skirball Institute of 
      Biomolecular Medicine, New York University, School of Medicine, New York, New 
      York, United States of America.
FAU - Schreiter, Eric
AU  - Schreiter E
AD  - Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, United 
      States of America.
FAU - Looger, Loren
AU  - Looger L
AD  - Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, United 
      States of America.
FAU - Neubert, Thomas
AU  - Neubert T
AD  - Department of Biochemistry and Molecular Pharmacology, Skirball Institute of 
      Biomolecular Medicine, New York University, School of Medicine, New York, New 
      York, United States of America.
FAU - Suh, Greg
AU  - Suh G
AD  - Department of Cell Biology, Skirball Institute of Biomolecular Medicine, New York 
      University, School of Medicine, New York, New York, United States of America.
LA  - eng
GR  - R01DC01279-01/DC/NIDCD NIH HHS/United States
GR  - P30 NS050276/NS/NINDS NIH HHS/United States
GR  - R01GM08946-01/GM/NIGMS NIH HHS/United States
GR  - R01 GM089746/GM/NIGMS NIH HHS/United States
GR  - S10RR027990/RR/NCRR NIH HHS/United States
GR  - S10 RR027990/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20150918
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Calmodulin)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 2.7.11.18 (Myosin-Light-Chain Kinase)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Calcium/metabolism
MH  - Calmodulin/*chemistry/genetics/metabolism
MH  - Color
MH  - Drosophila melanogaster
MH  - Fluorescence
MH  - Green Fluorescent Proteins/*chemistry/genetics/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Myosin-Light-Chain Kinase/*chemistry/genetics/metabolism
MH  - Peptide Fragments/chemistry/genetics/metabolism
MH  - *Photochemical Processes
MH  - Recombinant Fusion Proteins/*chemistry/genetics/metabolism
PMC - PMC4575167
COIS- Competing Interests: A patent application for this work has been filed, 
      Green-to-red photo-convertible fluorescent calcium indicator, application number 
      14/622,025. There are no further patents, products in development or marketed 
      products to declare. This does not alter the authors' adherence to all the PLOS 
      ONE policies on sharing data and materials, as detailed online in the guide for 
      authors.
EDAT- 2015/09/19 06:00
MHDA- 2016/06/09 06:00
PMCR- 2015/09/18
CRDT- 2015/09/19 06:00
PHST- 2015/02/16 00:00 [received]
PHST- 2015/08/25 00:00 [accepted]
PHST- 2015/09/19 06:00 [entrez]
PHST- 2015/09/19 06:00 [pubmed]
PHST- 2016/06/09 06:00 [medline]
PHST- 2015/09/18 00:00 [pmc-release]
AID - PONE-D-15-07287 [pii]
AID - 10.1371/journal.pone.0138127 [doi]
PST - epublish
SO  - PLoS One. 2015 Sep 18;10(9):e0138127. doi: 10.1371/journal.pone.0138127. 
      eCollection 2015.