Mini Chromosome and Fluorescent Proteins for Expression of Protein Complexes

Summary abstract The proteins encoded by the human genome may participate in as many as pairwise protein interactions and nearly every major process in a cell is carried out by assemblies of or more interlocking protein molecules Bruce Alberts Cell However there are no commercially available expression systems for regulated expression of multi subunit protein complexes There is an acute need for a mammalian expression system that can produce multi protein complexes is tightly regulated stable in prolonged culture and non integrated No reported expression system comes near to meeting all these needs Lucigen will develop and commercialize a stable episomal expression system with unbiased cloning ability tunable light inducible promoters improved fluorescent tags and the ability to express equivalent amounts of multiple proteins This system will facilitate direct experimental validation of the myriad protein complexes predicted by computational models as well as those that are already known e g ion channels multi subunit enzymes biochemical pathways This Phase II research will result in a novel mammalian expression vector light inducible promoters new fluorescent proteins for high throughput assays and stable cell lines that will greatly facilitate cloning expression and high throughput screening for numerous research and pharmaceutical applications The products developed in this proposal will be critical for functional analysis of the human protein interactome in normal and diseased states This knowledge will inform and confirm computational and in vitro models enabling critical advances in basic research and development of therapeutics