ArticleNature protocols2016
In vivo monitoring of cellular energy metabolism using SoNar, a highly responsive sensor for NAD(+)/NADH redox state.
Article in Nature protocols, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 81 papers.
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Who cites it
81 citing papers in PubMed, 165 citations in OpenAlex.
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- Data Augmentation Via Generative Adversarial Networks for Robust Prediction of NADH and pH from Limited Time-Resolved Fluorescence Spectra.Journal of fluorescence · 2026Article
- Mitochondrial uncoupling inhibits serine catabolismCancer biology & medicine · 2026Article
- Nonionotropic action of an acid-sensing ion channel inhibits leukemogenesis in the acidic bone marrow niche.The Journal of clinical investigation · 2025Article
- Nicotinamide Mononucleotide Enhances Boar Sperm Quality via Maintaining Mitochondrial Function During Liquid Storage.Animals : an open access journal from MDPI · 2025Article
- Restoring mitochondrial quantity and quality to reverse the Warburg effect and drive neuroblastoma differentiation.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- The Roles of Lactate and Lactylation in Diseases Related to Mitochondrial Dysfunction.International journal of molecular sciences · 2025Review
- Advances and trends for astaxanthin synthesis in Phaffia rhodozyma.Microbial cell factories · 2025Review
- Lactate activates trained immunity by fueling the tricarboxylic acid cycle and regulating histone lactylation.Nature communications · 2025Article
- Decreased mitochondrial NAD+ in WRN deficient cells links to dysfunctional proliferation.Aging · 2025Article
- Fast autofluorescence imaging to evaluate dynamic changes in cell metabolism.Journal of biomedical optics · 2024Article
- Elucidating the spatiotemporal dynamics of glucose metabolism with genetically encoded fluorescent biosensors.Cell reports methods · 2024Review
- Metabolic reprogramming in skeletal cell differentiation.Bone research · 2024Review
- Glial ferritin maintains neural stem cells via transporting iron required for self-renewal ineLife · 2024Article
- HES4 controls redox balance and supports pyrimidine synthesis and tumor growth.Nature structural & molecular biology · 2024Article
- RNAi screens identify HES4 as a regulator of redox balance supporting pyrimidine synthesis and tumor growth.Nature structural & molecular biology · 2024Article
21 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
Abstract
NADH and its oxidized form NAD(+) have a central role in energy metabolism, and their concentrations are often considered to be among the most important readouts of metabolic state. Here, we present a detailed protocol to image and monitor NAD(+)/NADH redox state in living cells and in vivo using a highly responsive, genetically encoded fluorescent sensor known as SoNar (sensor of NAD(H) redox). The chimeric SoNar protein was initially developed by inserting circularly permuted yellow fluorescent protein (cpYFP) into the NADH-binding domain of Rex protein from Thermus aquaticus (T-Rex). It functions by binding to either NAD(+) or NADH, thus inducing protein conformational changes that affect its fluorescent properties. We first describe steps for how to establish SoNar-expressing cells, and then discuss how to use the system to quantify the intracellular redox state. This approach is sensitive, accurate, simple and able to report subtle perturbations of various pathways of energy metabolism in real time. We also detail the application of SoNar to high-throughput chemical screening of candidate compounds targeting cell metabolism in a microplate-reader-based assay, along with in vivo fluorescence imaging of tumor xenografts expressing SoNar in mice. Typically, the approximate time frame for fluorescence imaging of SoNar is 30 min for living cells and 60 min for living mice. For high-throughput chemical screening in a 384-well-plate assay, the whole procedure generally takes no longer than 60 min to assess the effects of 380 compounds on cell metabolism.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.