In vitro
identification of novel nuclear receptor ligands
Human nuclear
receptors make up a family of approximately 50 different members. These
receptor proteins are expressed tissue- and cell-specifically and are
regulated by diverse stimuli including small molecules (ligands). Nuclear
receptors bind as monomers, heterodimers or homodimers to conserved DNA
response elements that are arranged in repeats in the promoter regions of
target genes. In vitro screening assays will be performed using nuclear
receptors overexpressed in cell line culture systems. Chemical libraries
will be used to treat these cell lines to determine whether new
receptor-modulating ligands may be identified and exploited for
therapeutic purposes. The proposed experiments will require the design of
mammalian expression plasmids and luciferase reporter vectors, and the
design and completion of nuclear receptor assays will teach students many
basic molecular biology techniques including DNA manipulation, polymerase
chain reaction (PCR), restriction endonuclease digestions and agarose gel
electrophoresis, UV/VIS spectrophotometry, mammalian cell line culture,
DNA transient transfection, gene reporter assays and luminescent
microplate assays. Additionally, students will learn to analyze and
interpret experimental data and reinforce understanding of statistical
analyses and the use of bioinformatics tools, as well as the careful
archiving of data and ability to modify experimental approaches based on
results. Skills learned in the laboratory will be demonstrated in routine
laboratory meetings and in the final SURF poster session. These skills
will further equip Fellows to continue their course of study at Masters or
PhD level and provide an early introduction to the scientific method; a
bonus will be the possibility of authorship on publications arising from
the proposed studies.
