alexa RNA-guided human genome engineering via Cas9.
Genetics

Genetics

Advancements in Genetic Engineering

Author(s): Mali P, Yang L, Esvelt KM, Aach J, Guell M,

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Abstract Bacteria 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.
This article was published in Science and referenced in Advancements in Genetic Engineering

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