A nanoparticle delivery system for CRISPR Cas based therapeutics

Abstract Applications of customizable nucleases such as CRISPR clustered regularly interspaced short palindromic repeats Cas CRISPR associated protein have enabled efficient and precise gene correction in vitro and hold promises for eventually achieving in vivo gene correction therapy However to apply CRISPR Cas in therapeutic settings several major challenges remain to be addressed i homologous recombination HR even with the help of Cas is still of low efficiency ii Cas is associated with off target effects and iii there is a lack of an efficient virus free system to deliver CRISPR Cas elements in vivo The present proposal focuses on the challenge of lack of an efficient non viral in vivo delivery system We recently developed novel hyperbranched polymers HPs with high nucleic acid binding affinity negligible cytotoxicity and achieved satisfactory delivery of microRNAs both in vitro and in vivo Here we propose to develop hyperbranched HP based system for effective delivery of Cas plasmid DNA pDNA In Phase I we will work to formulate and test new HPs for effectively packaging of Cas pDNA and evaluate the safety and efficacy of these HPs in vitro In Phase II we will expand the capability of HPs for Cas in vivo and apply HPs for Cas therapeutics in animal models Success of the proposed work will have significant impacts on basic and translational research and accelerate the development of Cas therapeutics Project Narrative One major challenge for CRISPR clustered regularly interspaced short palindromic repeats Cas CRISPR associated protein therapeutics is the lack of an efficient non viral in vivo delivery system The present work aims to develop a nanopolymer platform for delivery of Cas plasmid DNA both in vitro and in vivo Success of the proposed work will have significant impacts on basic and translational research and accelerate the development of Cas therapeutics