Mali, PrashantYang, LuhanEsvelt, Kevin M.Aach, JohnGüell Cargol, Marc, 1982-DiCarlo, James E.Norville, Julie E.Church, George M.2019-07-042019-07-042013Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE et al. RNA-guided human genome engineering via Cas9. Science. 2013 Feb 15;339(6121):823-6. DOI: 10.1126/science.12320330036-8075http://hdl.handle.net/10230/41936Bacteria and archaea have evolved adaptive immune defenses, termed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems, that use short RNA to direct degradation of foreign nucleic acids. Here, we engineer the type II bacterial CRISPR system to function with custom guide RNA (gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4% in induced pluripotent stem cells. We show that this process relies on CRISPR components; is sequence-specific; and, upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci. We also compute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons. Our results establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.application/pdfengThis is the author’s version of the work. It is posted here by permission of the AAAS for personal use, not for redistribution. The definitive version was published in Science on volume 339, number 6121, 2013, DOI: 10.1126/science.1232033Genoma humàEnginyeria genèticaRNAinfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1126/science.1232033info:eu-repo/semantics/openAccess