| AVS 65th International Symposium & Exhibition | |
| Biomaterial Interfaces Division | Tuesday Sessions |
| Session BI-TuP |
| Session: | Biomaterial Interfaces Division Poster Session |
| Presenter: | Telmo Diez, University of New Mexico |
| Authors: | T.D.P. Diez, University of New Mexico G.P. Lopez, University of New Mexico N.J. Carroll, University of New Mexico |
| Correspondent: | Click to Email |
Intrinsically disordered proteins (IDPs) are dynamic polypeptides used by eukaryotic cells in cell signaling, transcription, and chromatin remodeling functions are frequently employed in packaging and un-packaging of nucleic acids (NAs). Elastin-like polypeptides (ELPs) are biosynthetic biopolymers that have similar structural features to natural IDPs. Importantly, ELPs condense to form coacervates above a lower critical solution temperature (LCST). In this research, we focus on the combination of thermally responsive ELPs with natural nucleic acid binding domains to create promising responsive engineered protein constructs. In this study, we show how an ELP comprising nine positive charges from eight lysine interspersed within the chain is capable of interacting with NAs above its LCST. We characterize the amount of DNA captured by ELP and we use microfluidics to form aqueous microdroplets containing the ELP and fluorescent DNA to visualize DNA capture within ELP coacervate spheres via fluorescence microscopy. We characterize the thermodynamic binodal phase boundary (i.e. in the temperature-concentration dependent phase diagram) of the ELP[Office3] [#_msocom_3] /NA mixture to resolve the ELP volume fraction within the coacervate to predict the optimal temperature to maximize DNA capture. Finally, we combined this ELP with smaller RRM and RGG domains that bind nucleic acids found in natural FUS protein, a common NA binding protein that plays a role in genomic integrity.[Office4] [#_msocom_4] RRM is a 70 amino acid domain found to bind promiscuously to nucleic acids, and RGG is a 100 amino acid long domain rich on arginine and glycine found to be essential in the RRM interactions with nucleic acids. These studies have implications for, and yield insights into, the tailoring of engineered protein constructs that bind nucleic acids with predictable behavior and controlled release that could have many applications in gene therapy and other areas of bionanotechnology.