Research Interests:Aging appears to be the progressive loss of function through the deterioration of biological systems that control repair and maintenance. A multitude of factors are known to change with age, such as methylation patterns, enzyme activities, and production of inflammatory molecules. Clearly there are many changes in gene expression and changes in genes with age, but I believe primary authority over regulation of the rate of aging resides in the innate sequence of the genome. DNA encodes an organism’s biological processes, including its own repair and protection biomachinery. The rate of repair and protection and the rate of damage (whether endogenously or exogenously inflicted) together dictate the rate of deterioration of an organism. My long-standing interest is the aging process, and my goal at the Institute for Genome Sciences is to employ the power of modern genomic technology to elucidate genes associated with longevity and the genetic changes that contributed to the evolution of human longevity. There are widely divergent lifespans among primate species, yet it appears that genes that modulate aging have been conserved in sequence and function among divergent species over millions of years. Neoteny has been proposed as a mechanism that contributed to the rise of many human traits, including increased life span as compared to other primates. Comparative genomics will reveal the biosecrets that allow humans to age more slowly than other primates. An analysis that determines to what extent the genetic variants that modulate longevity are conserved across different species could have useful biomedical applications. Although aging has been slowed and healthy lifespan prolonged in many different animal models, much remains to be understood. Do the longer-lived species have a more stable genetic apparatus? Superior repair and protection mechanisms of cellular constituents? The comparative genomics approach will allow us to decode which regions of the genome are responsible for the preservation and extension of health in longer-lived species.
Lab Techniques and Equipment:• PCR. qPCR. Primer and Probe design. Gene synthesis. Cloning.
• DNA purification: mammalian genomic, phage, bacterial, plasmid.
• DNA modification: nucleases, endonucleases, ligase, kinase, phosphatase.
• Gel electrophoresis of DNA: agarose and polyacrylamide.
• FIGE (Field Inversion Gel Electrophoresis).
• TAFE (Transverse Alternating Field Electrophoresis).
• Site Directed Mutagenesis.
• Homologous recombination.
• Southern Blots.
• Library Construction.
• Mammalian tissue culture maintenance. Sterile technique.
• Mammalian clonal cell lines: Initiation, growth, splitting, cryogenic storage.
• Hybridoma cell line creation for production of monoclonal antibodies.
• Cell fractionation: isolation of microsomes, SER, RER.
• Isolation of cell nuclei, extraction of chromatin (rodent and primate tissue).
• Protein Purification Techniques.
• Column chromatography: Ion-exchange, affinity, and exclusion.
• Cyanogen bromide activation of sepharose 4B affinity gel.
• ELISA. Protein Detection and Quantitation. Enzyme assays.
• Peptide conjugation.
• IgG, IgM immunodetection; immunodiffusion test.
• SDS Polyacrylamide gel electrophoresis. Tricine gels.
• Western Blots.
• Fluorescence anisotropy measurements by steady state fluorimeter.
• Bacteria cells: Growth, induction, cryogenic storage.
• Purification of Lambda phage, T4 phage, M13.
• Phage T4 Expression-Packaging-Processing System to package and produce proteins.
• Production of proteins: by IPTG induction, heat-induction.
• Construction and production of chimera proteins.
• Phage Display.
Mills, R.E., Pittard, W.S., Mullaney, J.M., Farooq, U., Creasy, T.H., Mahurkar, A.A., Kemeza, D.M., Strassler, D.S., Ponting, C.P., Webber, C., and Devine, S.E. 2011. Natural genetic variation caused by small insertions and deletions in the human genome. Genome Res. 2011 Apr 25. [Epub ahead of print] PMID: 21460062