2016
DOI: 10.1038/nmeth.3764
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Apollo-NADP+: a spectrally tunable family of genetically encoded sensors for NADP+

Abstract: NADPH-dependent antioxidant pathways have a critical role in scavenging hydrogen peroxide (H2O2) produced by oxidative phosphorylation. Inadequate scavenging results in H2O2 accumulation and can cause disease. To measure NADPH/NADP(+) redox states, we explored genetically encoded sensors based on steady-state fluorescence anisotropy due to FRET (fluorescence resonance energy transfer) between homologous fluorescent proteins (homoFRET); we refer to these sensors as Apollo sensors. We created an Apollo sensor fo… Show more

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Cited by 108 publications
(107 citation statements)
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“…However, expression of WT NNT in J-islets did not restore the GRX1-roGFP2 response to glucose deprivation (Figure 2E). These results are compatible with recent data showing that the rise in cytosolic NADPH occurs between G0 and G5 [27], and indicate that the impact of NNT on cytosolic glutathione oxidation is negligible under control conditions.…”
Section: Resultssupporting
confidence: 92%
“…However, expression of WT NNT in J-islets did not restore the GRX1-roGFP2 response to glucose deprivation (Figure 2E). These results are compatible with recent data showing that the rise in cytosolic NADPH occurs between G0 and G5 [27], and indicate that the impact of NNT on cytosolic glutathione oxidation is negligible under control conditions.…”
Section: Resultssupporting
confidence: 92%
“…Dye based fluorescent probes have been designed for in vitro and in vivo imaging of glutathione in ultra-trace amount in mitochondria and platelet (Zhang et al, 2016 a,b). More recently, spectrally tunable genetically encoded sensor has been constructed for in vivo detection of NADP + (Cameron et al, 2016). Here, we report the construction, characterization and application of genetically encoded fluorescent nanosensor for non-invasive detection of the GB in living cells.…”
Section: Introductionmentioning
confidence: 99%
“…Therefore, more advanced methods need to be employed in the future to achieve chemical specificity and subcompartmental resolution. Technical possibilities include (i) the separation of NADH and NADPH autofluorescence using fluorescence lifetime imaging [24], (ii) the use of genetically encoded NADH [25] or NADþ/ NADH probes [26], and (iii) the use of the genetically encoded Apollo-NADP þ probe [27].…”
Section: Discussionmentioning
confidence: 99%