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An Expanded Palette of Genetically Encoded Ca2+ Indicators
Yongxin Zhao,1
Satoko Araki,2
Jiahui Wu,1
Takayuki Teramoto,3
Yu-Fen Chang,2
Masahiro Nakano,2
Ahmed S. Abdelfattah,1
Manabi Fujiwara,3
Takeshi Ishihara,3
Takeharu Nagai,2,4
Robert E. Campbell1,*
Abstract:
Engineered fluorescent protein (FP) chimeras that modulate their fluorescence in response to changes in calcium ion (Ca2+) concentration are powerful tools for visualizing intracellular signaling activity. However, despite a decade of availability, the palette of single FP-based Ca2+ indicators has remained limited to a single green hue. We have expanded this palette by developing blue, improved green, and red intensiometric indicators, as well as an emission ratiometric indicator with an 11,000% ratio change. This series enables improved single-color Ca2+ imaging in neurons and transgenic Caenorhabditis elegans. In HeLa cells, Ca2+ was imaged in three subcellular compartments, and, in conjunction with a cyan FP–yellow FP–based indicator, Ca2+ and adenosine 5'-triphosphate were simultaneously imaged. This palette of indicators paints the way to a colorful new era of Ca2+ imaging.
1 Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada. 2 Research Institute for Electronic Science, Hokkaido University, Kita 20, Nishi 10 Kita-ku, Sapporo 001-0020, Japan. 3 Department of Biology, Faculty of Sciences, Kyushu University, 6-10-1, Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan. 4 PRESTO, Japan Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo 102-0075, Japan.
* To whom correspondence should be addressed. E-mail: robert.e.campbell{at}ualberta.ca
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