(168w) Translating Polymeric Vehicles between Ribonucleoprotein and Plasmid DNA Cargoes: Do the Same Design Rules Apply?

Synthetic polymers are rapidly emerging as promising alternatives to viral delivery of therapeutic nucleic acid payloads such as DNA, RNA and ribonucleoproteins (RNP). Whether structure-function trends that are identified for a specific nucleic payload can be reapplied to other cargo types remains an unresolved question. In this work, we identified effective vehicles for plasmid DNA (pDNA) by screening a chemically diverse polymer library consisting of 43 unique structures. We discovered that a polymer previously identified for RNP delivery (P38) also emerged as the lead structure during screening studies for pDNA delivery, suggesting considerable overlap in the design rules for RNP and pDNA payloads. Through statistical learning tools we identified structural drivers for internalization, toxicity and transfection efficiency for both RNP and pDNA payloads and established that pDNA delivery is less reliant on hydrophobic interactions than RNPs. In addition to mediating transient transfection efficiently, P38 also co-delivered RNPs and pDNA donors to effect homology-directed repair (HDR) at higher rates than JetPEI. Our approach yielded a versatile transfection reagent that delivers multi-modal cargoes to accomplish challenging therapeutic objectives such as HDR.