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7208370 
Journal Article 
High-performance FRET biosensors for single-cell and in vivo lead detection 
Yang, DeM; Phan Nguyen Nhi Nguyen; Chang, YuFen; Fu, TF; Lin, CSea; Chiu, TaiYu; Huang, CC; Huang, HYi; Chung, MinWen; Lin, YuS; Manurung, RV; , 
2020 
Yes 
Biosensors and Bioelectronics
ISSN: 0956-5663
EISSN: 1873-4235 
ELSEVIER ADVANCED TECHNOLOGY 
OXFORD 
32892119 
WOS:000574928500003 
Forms of lead (Pb) have been insidiously invading human life for thousands of years without obvious signs of their considerable danger to human health. Blood lead level (BLL) is the routine measure used for diagnosing the degree of lead intoxication, although it is unclear whether there is any safe range of BLL. To develop a practical detection tool for living organisms, we engineered a genetically encoded fluorescence resonance energy transfer (FRET)-based Pb2+ biosensor, 'Met-lead 1.44 M1', with excellent performance. Met-lead 1.44 Ml has an apparent dissociation constant (K-d) of 25.97 nM, a detection limit (LOD) of 10 nM (2.0 ppb/0.2 mu g/dL), and an enhancement dynamic ratio of nearly similar to 5-fold upon Pb2+ binding. The 10 nM sensitivity of Met-lead 1.44 M1 is five times below the World Health Organization-permitted level of lead in tap water (10 ppb; WHO, 2017), and fifteen times lower than the maximum BLL for children (3 mu g/dL). We deployed Met-lead 1.44 M1 to measure Pb2+ concentrations in different living models, including two general human cell lines and one specific line, induced pluripotent stem cell (iPSC)-derived cardiomyocytes, as well as in widely used model species in plant (Arabidopsis thaliana) and animal (Drosophila melanogaster) research. Our results suggest that this new biosensor is suitable for lead toxicological research in vitro and in vivo, and will pave the way toward potential applications for both low BLL measures and rapid detection of environmental lead in its divalent form.