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