Richard Manderville

Instrumentation

Agilent HPLC with UV-vis/Fluorescence detection; BioAutomation MerMade 12 DNA Synthesizer; Cary 300-Bio UV-Vis spectrophotometer; Cary Eclipse and Edinburgh FS5 Spectrofluorometers.

Capabilities

Equipment allows the Manderville laboratory to synthesize modified DNA containing fluorescence DNA replacements. The modified DNA is purified using HPLC and then analyzed for DNA duplex stability and fluorescent performance of the dye using UV-Vis and fluorescence spectroscopies.

Education and Employment Background

Dr. Richard Manderville received his PhD from Queen’s University in 1992. He went on to work as a Postdoctoral Research Associate in BioOrganic Chemistry at the University of Virginia and an Associate Professor of Chemistry at Wake Forest University in Winston-Salem, North Carolina. He joined the University of Guelph in 2004 where he is currently a full professor of Chemistry and director of the Toxicology program.

Research Themes

Manderville’s research is focused on biochemical toxicology and biosensing applications.

1. Biochemical Toxicology. Manderville’s research in the area of biochemical toxicology focuses on the structural and biological impact of C8-aryl-2′-deoxyguanosine (C8-aryl-dG) adducts (addition products) that are a common lesion type and may contain N-, O- or C-aryl linkages (denoted N-, O- and C-linked adducts). The long-term goal of this project is to advance our fundamental understanding of how bulky C8-aryl-dG adducts containing various linkage types are processed in cells. To achieve this goal, synthetic organic chemistry is utilized to convert dG into various C8-aryl-dG adducts. The modified nucleoside is converted into a phosphoramidite and incorporated into oligonucleotide substrates using solid-phase DNA synthesis to afford adducted DNAs containing a single C8-aryl-dG adduct. Optical spectroscopies (UV-vis, fluorescence and circular dichroism) are then used to determine the impact of the lesion on duplex DNA. The adducted DNAs are also used as substrates in primer-extension assays with various DNA polymerase enzymes to determine the biological impact of the modified base. Students interested in molecular toxicology, organic and biochemistry, are encouraged to read some of our recent publications and inquire about possible projects in this area of biochemical toxicology.
2. Biosensing Applications. Fluorescence is one of the most powerful bioanalytical methods and fluorescent biosensors are designed to change their fluorescent intensities or wavelengths in response to physiological changes including pH, solvent polarity, viscosity, redox reactions, metal ions and apoptosis. This aspect of research focuses on the utility of internal fluorescent DNA bases to report molecular target binding by DNA aptamers. DNA aptamers are selected in vitro from random libraries for their ability to bind molecular targets with high affinity and specificity. Modification of DNA aptamers with internal fluorescent replacements can be used for diagnostics. The challenge is to select a fluorescent dye with sufficient brightness, chemical and photochemical stability, and emission sensitivity to target binding without perturbing aptamer affinity for the target.

Highlights

• NSERC Discovery Grant, 2004-present
• Director of Toxicology Program, University of Guelph, 2010-present
• NSERC Collaborative Research and Development Grant, 2017
• Ontario Centres of Excellence Grant, 2017
• Review member of NSERC Discovery Grants for Chemistry 2020