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Huiwang Ai

Huiwang Ai

Professor, Molecular Physiology and Biological Physics

Primary Location
1340 Jefferson Park Ave
Pinn Hall Rm. 4020, PO Box 800736
Charlottesville, VA 22908
Primary Phone Number
434.924.5665
Email
[email protected]
Website
https://med.virginia.edu/ai-lab/

Bio & Overview

The Ai lab specializes in protein engineering and employs interdisciplinary approaches, including biophysics, bioanalytical chemistry, chemical biology, synthetic biology, and optical imaging. One of our main areas of focus is the development of molecular biosensors that provide insights into cellular and organismal signaling. We employ state-of-the-art techniques such as directed evolution, fluorescence imaging, bioluminescence imaging, synthetic chemistry, genetic screening, and mass spectrometry to unravel complex biological pathways related to redox biology, metabolism, and neuronal activity. Furthermore, we explore the application of protein-based tools in live cells and rodent models to better understand pathophysiology and develop innovative therapies for conditions like diabetes, neurological disorders, and cancer. As an emerging direction, we are also working on the development of biosensors that can serve as self-wellness alert devices or diagnostics in resource-limited settings. List of Publications in Pubmed

Academic Information

Department
Molecular Physiology and Biological Physics
Academic Title
Associate Professor

Primary Education

  • BS, Chemistry, Tsinghua University, 2003
  • PhD, Chemical Biology, University of Alberta, 2008
  • Postdoc, Chemical Biology, The Scripps Research Institute, 2011

Other Appointments

  • Associate Professor, Chemistry
  • Associate Professor, Cell Biology

Other Affiliations

  • Cancer Center
  • Arts & Sciences
  • MSTP
  • BIMS

Research Interests

Molecular Biosensors; Spatiotemporal Regulation of Biological Signaling; Protein Engineering for Imaging, Diagnostics, and Therapeutics

Research Disciplines

  • Biochemistry
  • Biology
  • Biomedical Engineering
  • Biophysics
  • Biophysics & Structural Biology
  • Biotechnology
  • Cancer Biology
  • Cell and Developmental Biology
  • Chemistry
  • Metabolism
  • Molecular Biology
  • Molecular Pharmacology
  • Molecular Physiology and Biological Physics
  • Molecular and Cellular Physiology
  • Neuroscience
  • Physiology
  • Translational Science

Research Opportunities

  • Biochemistry
  • Biotechnology
  • Chemistry and Physics of Living Systems
  • Molecular and Systems Neuroscience
  • Physiological Basis of Human Disease

PhD Programs

  • Biophysics
  • Cell Biology
  • Physiology

Selected Publications

2024

Zhao, S., Xiong, Y., Sunnapu, R., Zhang, Y., Tian, X., & Ai, H. -W. (2024). Bioluminescence Imaging of Potassium Ion Using a Sensory Luciferin and an Engineered Luciferase. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 146(19), 13406-13416. doi:10.1021/jacs.4c02473

Tian, X., Zhang, Y., & Ai, H. -W. (2024). ATP-Independent Water-Soluble Luciferins Enable Non-Invasive High-Speed Video-Rate Bioluminescence Imaging of Mice.. bioRxiv. doi:10.1101/2024.04.30.591933

Zhao, S., Xiong, Y., Sunnapu, R., Zhang, Y., Tian, X., & Ai, H. -W. (2024). Bioluminescence Imaging of Potassium Ion Using a Sensory Luciferin and an Engineered Luciferase.. bioRxiv. doi:10.1101/2024.03.13.581057

Zhang, H., Tian, X., Zhang, J., & Ai, H. -W. (2024). Engineering and Characterization of 3-Aminotyrosine-Derived Red Fluorescent Variants of Circularly Permutated Green Fluorescent Protein. BIOSENSORS-BASEL, 14(1). doi:10.3390/bios14010054

2023

Zhang, J., Li, Z., Pang, Y., Fan, Y., & Ai, H. -W. (2023). Genetically Encoded Boronolectin as a Specific Red Fluorescent UDP-GlcNAc Biosensor. ACS SENSORS. doi:10.1021/acssensors.3c00409

Chen, C., Zhang, H., Zhang, J., Ai, H. -W., & Fang, C. (2023). Structural origin and rational development of bright red noncanonical variants of green fluorescent protein. PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 25(23), 15624-15634. doi:10.1039/d3cp01315d

Pang, Y., Huang, M., Fan, Y., Yeh, H. -W., Xiong, Y., Ng, H. L., & Ai, H. -W. (2023). Development, Characterization, and Structural Analysis of a Genetically Encoded Red Fluorescent Peroxynitrite Biosensor. ACS CHEMICAL BIOLOGY, 18(6), 1388-1397. doi:10.1021/acschembio.3c00139

Zhang, J., Li, Z., Pang, Y., Fan, Y., & Ai, H. -W. (2023). Genetically Encoded Boronolectin as a Specific Red Fluorescent UDP-GlcNAc Biosensor.. bioRxiv. doi:10.1101/2023.03.01.530644

Wu, T., Kumar, M., Zhang, J., Zhao, S., Drobizhev, M., McCollum, M., . . . Ai, H. -W. (2023). A genetically encoded far-red fluorescent indicator for imaging synaptically released Zn2.. Science advances, 9(9), eadd2058. doi:10.1126/sciadv.add2058

2022

Dulin, H., Hendricks, N., Xu, D., Gao, L., Wuang, K., Ai, H., & Hai, R. (2022). Impact of Protein Nitration on Influenza Virus Infectivity and Immunogenicity.. Microbiology spectrum, 10(6), e0190222. doi:10.1128/spectrum.01902-22

Xiong, Y., Zhang, Y., Li, Z., Reza, M. S., Li, X., Tian, X., & Ai, H. -W. (2022). Engineered Amber-Emitting Nano Luciferase and Its Use for Immunobioluminescence Imaging In Vivo . JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 144(31), 14101-14111. doi:10.1021/jacs.2c02320

Tian, X., Zhang, Y., Li, X., Xiong, Y., Wu, T., & Ai, H. -W. (2022). A luciferase prosubstrate and a red bioluminescent calcium indicator for imaging neuronal activity in mice. NATURE COMMUNICATIONS, 13(1). doi:10.1038/s41467-022-31673-x

Li, X., Zhang, Y., & Ai, H. -W. (2022). Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in Aβ42-Mediated Neurotoxicity. ACS SENSORS, 7(3), 722-729. doi:10.1021/acssensors.1c01381

Guo, Z., Parakra, R. D., Xiong, Y., Johnston, W. A., Walden, P., Edwardraja, S., . . . Alexandrov, K. (2022). Engineering and exploiting synthetic allostery of NanoLuc luciferase.. Nature communications, 13(1), 789. doi:10.1038/s41467-022-28425-2

Pang, Y., Zhang, H., & Ai, H. -W. (2022). Improved Red Fluorescent Redox Indicators for Monitoring Cytosolic and Mitochondrial Thioredoxin Redox Dynamics. BIOCHEMISTRY, 61(5), 1-8. doi:10.1021/acs.biochem.1c00634

Yu, X., Xing, Y., Zhang, Y., Zhang, P., He, Y., Ghamsari, F., . . . Oberholzer, J. (2022). Smartphone-microfluidic fluorescence imaging system for studying islet physiology. FRONTIERS IN ENDOCRINOLOGY, 13. doi:10.3389/fendo.2022.1039912

2021

Wu, T., Pang, Y., & Ai, H. -W. (2021). Circularly Permuted Far-Red Fluorescent Proteins. BIOSENSORS-BASEL, 11(11). doi:10.3390/bios11110438

Erdogan, Y. C., Altun, H. Y., Secilmis, M., Ata, B. N., Sevimli, G., Cokluk, Z., . . . Eroglu, E. (2021). Complexities of the chemogenetic toolkit: Differential mDAAO activation by d-amino substrates and subcellular targeting.. Free radical biology & medicine, 177, 132-142. doi:10.1016/j.freeradbiomed.2021.10.023

Li, Z., Zhang, J., & Ai, H. -W. (2021). Genetically Encoded Green Fluorescent Biosensors for Monitoring UDP-GlcNAc in Live Cells. ACS CENTRAL SCIENCE, 7(10), 1763-1770. doi:10.1021/acscentsci.1c00745

Pang, Y., Zhang, H., & Ai, H. -W. (2021). Genetically Encoded Fluorescent Redox Indicators for Unveiling Redox Signaling and Oxidative Toxicity. CHEMICAL RESEARCH IN TOXICOLOGY, 34(8), 1826-1845. doi:10.1021/acs.chemrestox.1c00149

Chen, Z., Zhang, S., Li, X., & Ai, H. -W. (2021). A high-performance genetically encoded fluorescent biosensor for imaging physiological peroxynitrite. CELL CHEMICAL BIOLOGY, 28(11), 1542-+. doi:10.1016/j.chembiol.2021.01.013

Yu, X., Zhang, P., He, Y., Lin, E., Ai, H., Ramasubramanian, M. K., . . . Oberholzer, J. (2021). A Smartphone-Fluidic Digital Imaging Analysis System for Pancreatic Islet Mass Quantification. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, 9. doi:10.3389/fbioe.2021.692686

2020

Zhang, S., & Ai, H. -W. (2020). A general strategy to red-shift green fluorescent protein-based biosensors. NATURE CHEMICAL BIOLOGY, 16(12), 1434-+. doi:10.1038/s41589-020-0641-7

2019

Chen, M., Zhang, S., Xing, Y., Li, X., He, Y., Wang, Y., . . . Ai, H. -W. (2019). Genetically Encoded, Photostable Indicators to Image Dynamic Zn2+ Secretion of Pancreatic Islets. ANALYTICAL CHEMISTRY, 91(19), 12212-12219. doi:10.1021/acs.analchem.9b01802

Achimovich, A. M., Ai, H., & Gahlmann, A. (2019). Enabling technologies in super-resolution fluorescence microscopy: reporters, labeling, and methods of measurement. CURRENT OPINION IN STRUCTURAL BIOLOGY, 58, 224-232. doi:10.1016/j.sbi.2019.05.001

Youssef, S., Zhang, S., & Ai, H. -W. (2019). A Genetically Encoded, Ratiometric Fluorescent Biosensor for Hydrogen Sulfide. ACS SENSORS, 4(6), 1626-1632. doi:10.1021/acssensors.9b00400

Xiong, Y., Tian, X., & Ai, H. -W. (2019). Molecular Tools to Generate Reactive Oxygen Species in Biological Systems. BIOCONJUGATE CHEMISTRY, 30(5), 1297-1303. doi:10.1021/acs.bioconjchem.9b00191

Yeh, H. -W., Xiong, Y., Wu, T., Chen, M., Ji, A., Li, X., & Ai, H. -W. (2019). ATP-Independent Bioluminescent Reporter Variants To Improve in Vivo Imaging. ACS CHEMICAL BIOLOGY, 14(5), 959-965. doi:10.1021/acschembio.9b00150

Yeh, H. -W., Wu, T., Chen, M., & Ai, H. -W. (2019). Identification of Factors Complicating Bioluminescence Imaging. BIOCHEMISTRY, 58(12), 1689-1697. doi:10.1021/acs.biochem.8b01303

Yeh, H. -W., & Ai, H. -W. (2019). Development and Applications of Bioluminescent and Chemiluminescent Reporters and Biosensors. ANNUAL REVIEW OF ANALYTICAL CHEMISTRY, VOL 12, 12, 129-150. doi:10.1146/annurev-anchem-061318-115027

2018

Ji, A., Fan, Y., Ren, W., Zhang, S., & Ai, H. -W. (2018). A Sensitive Near-Infrared Fluorescent Sensor for Mitochondrial Hydrogen Sulfide. ACS SENSORS, 3(5), 992-997. doi:10.1021/acssensors.8b00142

Gao, L., Tan, X. -F., Zhang, S., Wu, T., Zhang, Z. -M., Ai, H. -W., & Song, J. (2018). An Intramolecular Interaction of UHRF1 Reveals Dual Control for Its Histone Association. STRUCTURE, 26(2), 304-+. doi:10.1016/j.str.2017.12.016

2017

Yeh, H. -W., Karmach, O., Ji, A., Carter, D., Martins-Green, M. M., & Ai, H. -W. (2017). Red-shifted luciferase-luciferin pairs for enhanced bioluminescence imaging. NATURE METHODS, 14(10), 971-+. doi:10.1038/NMETH.4400

Fan, Y., Makar, M., Wang, M. X., & Ai, H. -W. (2017). Monitoring thioredoxin redox with a genetically encoded red fluorescent biosensor. NATURE CHEMICAL BIOLOGY, 13(9), 1045-+. doi:10.1038/NCHEMBIO.2417

Chen, Z., Truong, T. M., & Ai, H. -W. (2017). Illuminating Brain Activities with Fluorescent Protein-Based Biosensors. CHEMOSENSORS, 5(4). doi:10.3390/chemosensors5040032

2016

Chen, Z., & Ai, H. -W. (2016). Single Fluorescent Protein-Based Indicators for Zinc Ion (Zn2+). ANALYTICAL CHEMISTRY, 88(18), 9029-9036. doi:10.1021/acs.analchem.6b01653

Youssef, S., Ren, W., & Ai, H. -W. (2016). A Genetically Encoded FRET Sensor for Hypoxia and Prolyl Hydroxylases. ACS CHEMICAL BIOLOGY, 11(9), 2492-2498. doi:10.1021/acschembio.6b00330

Chen, Z. -J., Tian, Z., Kallio, K., Oleson, A. L., Ji, A., Borchardt, D., . . . Ai, H. -W. (2016). The N-B Interaction through a Water Bridge: Understanding the Chemoselectivity of a Fluorescent Protein Based Probe for Peroxynitrite. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 138(14), 4900-4907. doi:10.1021/jacs.6b01285

Fan, Y., & Ai, H. -W. (2016). Development of redox-sensitive red fluorescent proteins for imaging redox dynamics in cellular compartments (vol 408, pg 2901, 2016). ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 408(11), 3043. doi:10.1007/s00216-016-9365-7

Fan, Y., & Ai, H. -W. (2016). Development of redox-sensitive red fluorescent proteins for imaging redox dynamics in cellular compartments. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 408(11), 2901-2911. doi:10.1007/s00216-015-9280-3

Ren, W., Ji, A., Karmach, O., Carter, D. G., Martins-Green, M. M., & Ai, H. -W. (2016). A membrane-activatable near-infrared fluorescent probe with ultra-photostability for mitochondrial membrane potentials. ANALYST, 141(12), 3679-3685. doi:10.1039/c5an01860a

2015

Ren, W., Truong, T. M., & Ai, H. -W. (2015). Study of the Binding Energies between Unnatural Amino Acids and Engineered Orthogonal Tyrosyl-tRNA Synthetases. SCIENTIFIC REPORTS, 5. doi:10.1038/srep12632

Ren, W., Ji, A., Wang, M. X., & Ai, H. -W. (2015). Expanding the Genetic Code for a Dinitrophenyl Hapten. CHEMBIOCHEM, 16(14), 2007-2010. doi:10.1002/cbic.201500204

Fan, Y., Chen, Z., & Ai, H. -W. (2015). Monitoring Redox Dynamics in Living Cells with a Redox-Sensitive Red Fluorescent Protein. ANALYTICAL CHEMISTRY, 87(5), 2802-2810. doi:10.1021/ac5041988

Ren, W., Ji, A., & Ai, H. -W. (2015). Light Activation of Protein Splicing with a Photocaged Fast Intein. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 137(6), 2155-2158. doi:10.1021/ja508597d

2014

Ai, H. -W. (2015). Fluorescent-protein-based probes: general principles and practices. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 407(1), 9-15. doi:10.1007/s00216-014-8236-3

Ai, H. -W. (2014). Fluorescent Sensors for Biological Applications. SENSORS, 14(9), 17829-17831. doi:10.3390/s140917829

Chen, Z. -J., & Ai, H. -W. (2014). A Highly Responsive and Selective Fluorescent Probe for Imaging Physiological Hydrogen Sulfide. BIOCHEMISTRY, 53(37), 5966-5974. doi:10.1021/bi500830d

Ai, H. -W., Baird, M. A., Shen, Y., Davidson, M. W., & Campbell, R. E. (2014). Engineering and characterizing monomeric fluorescent proteins for live-cell imaging applications. NATURE PROTOCOLS, 9(4), 910-928. doi:10.1038/nprot.2014.054

Kang, M., Light, K., Ai, H. -W., Shen, W., Kim, C. H., Chen, P. R., . . . Schultz, P. G. (2014). Evolution of Iron(II)-Finger Peptides by Using a Bipyridyl Amino Acid. CHEMBIOCHEM, 15(6), 822-825. doi:10.1002/cbic.201300727

Ji, A., Ren, W., & Ai, H. -W. (2014). A highly efficient oxidative condensation reaction for selective protein conjugation. CHEMICAL COMMUNICATIONS, 50(56), 7469-7472. doi:10.1039/c4cc01551g

2013

Ren, W., & Ai, H. -W. (2013). Genetically Encoded Fluorescent Redox Probes. SENSORS, 13(11), 15422-15433. doi:10.3390/s131115422

Chen, Z. -J., Ren, W., Wright, Q. E., & Ai, H. -W. (2013). Genetically Encoded Fluorescent Probe for the Selective Detection of Peroxynitrite. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 135(40), 14940-14943. doi:10.1021/ja408011q

Chatterjee, A., Xiao, H., Bollong, M., Ai, H. -W., & Schultz, P. G. (2013). Efficient viral delivery system for unnatural amino acid mutagenesis in mammalian cells. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(29), 11803-11808. doi:10.1073/pnas.1309584110

2012

Chen, S., Chen, Z. -J., Ren, W., & Ai, H. -W. (2012). Reaction-Based Genetically Encoded Fluorescent Hydrogen Sulfide Sensors. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 134(23), 9589-9592. doi:10.1021/ja303261d

Ai, H. -W. (2012). Biochemical analysis with the expanded genetic lexicon. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 403(8), 2089-2102. doi:10.1007/s00216-012-5784-2

2011

Ding, Y., Ai, H. -W., Hoi, H., & Campbell, R. E. (2011). Forster Resonance Energy Transfer-Based Biosensors for Multiparameter Ratiometric Imaging of Ca2+ Dynamics and Caspase-3 Activity in Single Cells. ANALYTICAL CHEMISTRY, 83(24), 9687-9693. doi:10.1021/ac202595g

Ai, H. -W., Shen, W., Sagi, A., Chen, P. R., & Schultz, P. G. (2011). Probing Protein-Protein Interactions with a Genetically Encoded Photo-crosslinking Amino Acid. CHEMBIOCHEM, 12(12), 1854-1857. doi:10.1002/cbic.201100194

2010

Ai, H. -W., Lee, J. W., & Schultz, P. G. (2010). A method to site-specifically introduce methyllysine into proteins in E. coli. CHEMICAL COMMUNICATIONS, 46(30), 5506-5508. doi:10.1039/c0cc00108b

Ai, H. -W., Shen, W., Brustad, E., & Schultz, P. G. (2010). Genetically Encoded Alkenes in Yeast. ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 49(5), 935-937. doi:10.1002/anie.200905590