title: Genome surgery using Cas9 ribonucleoproteins for the treatment of age-related macular degeneration.
abstract: RNA-guided genome surgery using CRISPR-Cas9 nucleases has shown promise for the treatment of diverse genetic diseases. Yet, the potential of such nucleases for therapeutic applications in nongenetic diseases is largely unexplored. Here, we focus on age-related macular degeneration (AMD), a leading cause of blindness in adults, which is associated with retinal overexpression of, rather than mutations in, theVEGFAgene. Subretinal injection of preassembled,Vegfagene-specific Cas9 ribonucleoproteins (RNPs) into the adult mouse eye gave rise to mutagenesis at the target site in the retinal pigment epithelium. Furthermore, Cas9 RNPs effectively reduced the area of laser-induced choroidal neovascularization (CNV) in a mouse model of AMD. Genome-wide profiling of Cas9 off-target effects via Digenome-seq showed that off-target mutations were rarely induced in the human genome. Because Cas9 RNPs can function immediately after in vivo delivery and are rapidly degraded by endogenous proteases, their activities are unlikely to be hampered by antibody- and cell-mediated adaptive immune systems. Our results demonstrate that in vivo genome editing with Cas9 RNPs has the potential for the local treatment for nongenetic degenerative diseases, expanding the scope of RNA-guided genome surgery to a new dimension.
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.
authors.forename: Kyoungmi
authors.initials: K
authors.lastname: Kim
authors.affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.|FARB Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03082, Republic of Korea.
authors.forename: Sung Wook
authors.initials: SW
authors.lastname: Park
authors.affiliation: FARB Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03082, Republic of Korea.
authors.forename: Jin Hyoung
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.|Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Daesik
authors.initials: D
authors.lastname: Kim
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Koo
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.|Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Kwang-Eun
authors.initials: KE
authors.lastname: Kim
authors.affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.|FARB Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03082, Republic of Korea.|Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.
authors.forename: Jeong Hun
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Seoul 08826, Republic of Korea.|Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D000076987:CRISPR-Associated Protein 9; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism*; D000072669:Gene Editing / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008268:Macular Degeneration / Q000628:therapy*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D059748:Proteolysis; D012160:Retina / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D012261:Ribonucleoproteins; D042461:Vascular Endothelial Growth Factor A; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases