(638a) Solid Lipid Nanoparticles Based Protein Delivery Platform for Effective Non-Viral Gene Editing

Therapeutics based on proteins, RNA and DNA have tremendous therapeutic potential but have been challenging to develop due to delivery problems. Recently, CRISPR/Cas9-based therapeutics show great potentials to revolutionize the treatment of genetic diseases. However, the lack of safe and efficient delivery method is still a big challenge for its clinical translation. We have developed a novel solid lipid nanoparticles based delivery platform which can deliver Cas9 ribonucleoprotein (RNP) and donor DNA into wide variety of type for gene knockdown, DNA mutation correction and gene knockin. This delivery system shows five times increase of NHEJ based gene knockdown efficiency in HEK 293T cells and three times increase in primary myoblast cells compared with commercially available lipofectamine 2000 based formulation. Also, this delivery system shows efficient knockdown of pcsk9 gene in liver in vivo by intravenous injection, which significantly reduced pcsk9 and cholesterol level in the serum. Meanwhile, by delivering of Cas9 RNP and single strand donor DNA together, this delivery system can efficiently correct the DNA mutation that causes Duchenne muscular dystrophy in mice via local injection, with minimal off-target DNA damage. More importantly, this delivery method can knock in complete cytokine genes into tumor cells which have great potential for cancer immunotherapy.