Structure-property study for dendronized polymer vectors for CRISPR delivery

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Nontechnical Summary:This project develops a new molecular delivery system for enabling the CRISPR-based genome editing technology. CRISPR-based genome editing technology can precisely add, delete, and “correct” genes in vitro or in vivo; therefore, it holds tremendous potential for therapeutic applications and is actively being pursued for addressing a wide range of human diseases ranging from hereditary diseases, cancer, to diabetes. However, one major obstacle to implementing this technology is the lack of safe and efficient delivery systems that can effectively transport the required molecular machinery into cells to execute genome editing. To address this challenge, this project proposes the design, synthesis, and investigation of a series of delivery molecules made from natural amino acids and peptides. These compounds will be evaluated for their efficiency and safety for delivering two pieces of RNAs into cells for performing CRISPR-based genome editing. Structure-function studies will shed light on important design principles for effective molecular carriers. The proposed interdisciplinary research activities also provide excellent trainings to students at various levels, ranging from K-12, undergraduate, to graduate students, especially women, underrepresented minority students, and those from institutes with limited research opportunities.Technical Summary:The goal of this project is to develop a new synthetic delivery platform for RNA-based CRISPR and conduct fundamental structure-property studies on the new delivery system to gain critical insights for the design of new gene delivery vectors. CRISPR/Cas9 gene editing offers a tremendous potential for therapeutic applications and is actively being pursued for addressing a wide range of human diseases. However, one major obstacle to implementing CRISPR-mediated genome editing is the lack of safe and efficient delivery vehicles. The PI lab recently developed an innovative design of bioreducible dendronized polypeptides (BDPs) that show high efficiency for co-delivery of Cas9 mRNA and gRNAs. It is proposed that further investigation of this system will generate a new platform delivery system that can effectively and safely co-deliver CRISPR/Cas9 mRNA and gRNA into various cells. Furthermore, capitalizing on BDP’s well-defined molecular structure, we propose to conduct fundamental structure-property studies on the BDP system with the aim to gain critical insights for the design of new gene delivery vectors. If successful, the proposed study will provide a new, general, and biocompatible delivery system for CRISPR technology and offer critical insights for guiding the design of new gene delivery systems. The research and broad impact activities will provide holistic trainings to students at various levels, ranging from K-12, undergraduate, to graduate students, especially women, underrepresented minority students.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.