@article{Kim01062014,
author = {Kim, Sojung and Kim, Daesik and Cho, Seung Woo and Kim, Jungeun and Kim, Jin-Soo}, 
title = {Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins},
volume = {24}, 
number = {6}, 
pages = {1012-1019}, 
year = {2014}, 
doi = {10.1101/gr.171322.113}, 
abstract ={RNA-guided engineered nucleases (RGENs) derived from the prokaryotic adaptive immune system known as CRISPR (clustered, regularly interspaced, short palindromic repeat)/Cas (CRISPR-associated) enable genome editing in human cell lines, animals, and plants, but are limited by off-target effects and unwanted integration of DNA segments derived from plasmids encoding Cas9 and guide RNA at both on-target and off-target sites in the genome. Here, we deliver purified recombinant Cas9 protein and guide RNA into cultured human cells including hard-to-transfect fibroblasts and pluripotent stem cells. RGEN ribonucleoproteins (RNPs) induce site-specific mutations at frequencies of up to 79%, while reducing off-target mutations associated with plasmid transfection at off-target sites that differ by one or two nucleotides from on-target sites. RGEN RNPs cleave chromosomal DNA almost immediately after delivery and are degraded rapidly in cells, reducing off-target effects. Furthermore, RNP delivery is less stressful to human embryonic stem cells, producing at least twofold more colonies than does plasmid transfection.}, 
URL = {http://genome.cshlp.org/content/24/6/1012.abstract}, 
eprint = {http://genome.cshlp.org/content/24/6/1012.full.pdf+html}, 
journal = {Genome Research} 
}