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ARS Home » Plains Area » Houston, Texas » Children's Nutrition Research Center » Research » Publications at this Location » Publication #331554
Title: Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles

Author
item WANG, MING - Tufts University
item ZURIS, JOHN - Harvard University
item MENG, FANTAO - Children'S Nutrition Research Center (CNRC)
item REES, HELEN - Harvard University
item SUN, SHUO - Tufts University
item DENG, PING - Tufts University
item HAN, YONG - Children'S Nutrition Research Center (CNRC)
item GAO, XUE - Harvard University
item POULI, DIMITRA - Tufts University
item WU, QI - Children'S Nutrition Research Center (CNRC)
item GEORGAKOUDI, IRENE - Tufts University
item LIU, DAVID - Harvard University
item XU, QIAOBING - Tufts University

Submitted to: Proceedings of the National Academy of Sciences (PNAS)
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/2/2016
Publication Date: 8/1/2016
Citation: Wang, M., Zuris, J., Meng, F., Rees, H., Sun, S., Deng, P., Han, Y., Gao, X., Pouli, D., Wu, Q., Georgakoudi, I., Liu, D.R., Xu, Q. 2016. Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles. Proceedings of the National Academy of Sciences. 113(11):2868-2873.

Interpretive Summary: There is high demand in adopting a new genome-editing system to the neuroscience field. This report showed that a novel class of nanoparticles can effectively deliver gene-editing materials into desirable brain areas. This new nanotechnology warrants safe and efficient genetic analysis in the brain and may be used as a method in future research studies.

Technical Abstract: A central challenge to the development of protein-based therapeutics is the inefficiency of delivery of protein cargo across the mammalian cell membrane, including escape from endosomes. Here we report that combining bioreducible lipid nanoparticles with negatively supercharged Cre recombinase or anionic Cas9:single-guide (sg)RNA complexes drives the electrostatic assembly of nanoparticles that mediate potent protein delivery and genome editing. These bioreducible lipids efficiently deliver protein cargo into cells, facilitate the escape of protein from endosomes in response to the reductive intracellular environment, and direct protein to its intracellular target sites. The delivery of supercharged Cre protein and Cas9:sgRNA complexed with bioreducible lipids into cultured human cells enables gene recombination and genome editing with efficiencies greater than 70%. In addition, we demonstrate that these lipids are effective for functional protein delivery into mouse brain for gene recombination in vivo. Therefore, the integration of this bioreducible lipid platform with protein engineering has the potential to advance the therapeutic relevance of protein-based genome editing.