Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting

	Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting
	Bajar, Bryce T.; Wang, Emily S.; Lam, Amy J.; Kim, Bongjae B.; Jacobs, Conor L.; Howe, Elizabeth S.; Davidson, Michael W.; Lin, Michael Z.; Chu, Jun
刊名	SCIENTIFIC REPORTS
	2016
英文摘要	Many genetically encoded biosensors use Forster resonance energy transfer (FRET) to dynamically report biomolecular activities. While pairs of cyan and yellow fluorescent proteins (FPs) are most commonly used as FRET partner fluorophores, respectively, green and red FPs offer distinct advantages for FRET, such as greater spectral separation, less phototoxicity, and lower autofluorescence. We previously developed thegreen-red FRET pair Clover and mRuby2, which improves responsiveness in intramolecular FRET reporters with different designs. Here we report the engineering of brighter and more photostable variants, mClover3 and mRuby3. mClover3 improves photostability by 60% and mRuby3 by 200% over the previous generationof fluorophores. Notably, mRuby3 is also 35% brighter than mRuby2, making it both the brightest and most photostable monomeric red FP yet characterized. Furthermore, we developed a standardized methodologyfor assessing FP performance in mammalian cells as stand-alone markers and as FRET partners. We found that mClover3 or mRuby3 expression in mammalian cells provides the highest fluorescence signals of all jellyfish GFP or coral RFP derivatives, respectively. Finally, using mClover3 and mRuby3, we engineered an improved version of the CaMKII alpha reporter Camuia with a larger response amplitude.
收录类别	SCI
原文出处	http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4754705/
语种	英语
内容类型	期刊论文
源URL	[http://ir.siat.ac.cn:8080/handle/172644/10497]
专题	深圳先进技术研究院_医工所
作者单位	SCIENTIFIC REPORTS
推荐引用方式 GB/T 7714	Bajar, Bryce T.,Wang, Emily S.,Lam, Amy J.,et al. Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting[J]. SCIENTIFIC REPORTS,2016.
APA	Bajar, Bryce T..,Wang, Emily S..,Lam, Amy J..,Kim, Bongjae B..,Jacobs, Conor L..,...&Chu, Jun.(2016).Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.SCIENTIFIC REPORTS.
MLA	Bajar, Bryce T.,et al."Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting".SCIENTIFIC REPORTS (2016).