Growth of Hybrid Polymeric Nanostructures for Enzyme-Free Amplified Protein Imaging

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Proteins play essential roles in the body. However, because they have typically are found in low concentration, they can be very challenging to detect. Overcoming this challenge will not advance basic research and greatly improve the quality of diagnosis and the discovery of new therapeutics. The objective of this project is to develop a novel biosensing nanotechnology to amplify the signals of target proteins. The proteins that cannot be detected using traditional methods would be detected with this new technology. Students at different levels will be trained through hands-on research experience. Nationwide underrepresented students through diverse outreach activities at Penn State will be recruited to this program. The new knowledge to be generated through this program will be integrated into the classes offered to undergraduate and graduate students. Moreover, the knowledge will be broadly disseminated through high-profile journal publications and conference presentations. Protein examination is pivotal to virtually all fields related to biomedical sciences and clinical diagnostics as well. However, existing methods for in situ protein imaging and sensing often suffer from problems such as weak signal intensity, pigment/nanoparticle deposition and slow diffusion of bulky agents. The objective of this project is to explore a novel method for enzyme-free signal amplification in protein imaging via in situ growth of biomolecular nanostructures. The success of exploring this new method will address the critical unmet needs faced by existing methods for protein detection. In the proposed new method, each protein target will be represented by a supramolecular imaging nanostructure that carries a large number of fluorophores. Notably, the nanostructure will be grown in situ for imaging without the need of using any enzyme. Thus, it will not only provide signal amplification but also avoid potential enzyme- or nanoparticle-associated problems. The proposed research is designed with three tasks: 1) to understand how the compositions and structures of DNA molecules determine the formation of DNA nanostructures; 2) to synthesize and characterize hybrid DNA-based supramolecular nanostructures; and 3) to evaluate the effectiveness of this method in protein imaging. The accomplishment of this project will make broad impacts. Firstly, it will advance the methodology of protein sensing, benefiting a variety of biological and biomedical research areas that require highly sensitive protein imaging. Secondly, the fundamental understanding of DNA polymerization will provide a knowledge basis for the development of various DNA-based biosensors and materials. Thirdly, as clinical biopsies often have very small volumes, this enzyme-free signal amplification imaging method also holds great potential of improving clinical diagnostics. The broad impacts will be further strengthened by the development of human resources at different levels with active recruitment of nationwide underrepresented students through a diverse array of Penn State outreach programs, the publication of papers in high-profile journals and the dissemination of knowledge through invited seminars and conference presentations.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.