A nanoparticle delivery system for CRISPR/Cas9 based therapeutics

Abstract Applications of customizable nucleases such as CRISPRclustered regularly interspaced short palindromic repeatsCasCRISPR associated proteinhave enabled efficient and precise gene correction in vitroand hold promises for eventually achieving in vivo gene correction therapyHoweverto apply CRISPR Casin therapeutic settingsseveral major challenges remain to be addressedihomologous recombinationHReven with the help of Casis still of low efficiencyiiCasis associated with off target effectsandiiithere is a lack of an efficient virus free system to deliver CRISPR Caselements in vivoThe present proposal focuses on the challenge of lack of an efficient non viral in vivo delivery systemWe recently developed novel hyperbranched polymersHPswith high nucleic acid binding affinitynegligible cytotoxicityand achieved satisfactory delivery of microRNAs both in vitro and in vivoHere we propose to develop hyperbranched HPbased system for effective delivery of Casplasmid DNApDNAIn Phase Iwe will work to formulate and test new HPs for effectively packaging of CaspDNAand evaluate the safety and efficacy of these HPs in vitroIn Phase IIwe will expand the capability of HPs for Casin vivoand apply HPs for Castherapeutics in animal modelsSuccess of the proposed work will have significant impacts on basic and translational research and accelerate the development of Castherapeutics Project Narrative One major challenge for CRISPRclustered regularly interspaced short palindromic repeatsCasCRISPR associated proteintherapeutics is the lack of an efficient non viral in vivo delivery systemThe present work aims to develop a nanopolymer platform for delivery of Casplasmid DNA both in vitro and in vivoSuccess of the proposed work will have significant impacts on basic and translational researchand accelerate the development of Castherapeutics