Daesik Kim

TL;DR: Delivery of purified recombinant Cas9 protein and guide RNA into cultured human cells including hard-to-transfect fibroblasts and pluripotent stem cells is delivered and 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.

TL;DR: Digenome-seq is a robust, sensitive, unbiased and cost-effective method for profiling genome-wide off-target effects of programmable nucleases including Cas9, and shows that Cas9 off- target effects can be avoided by replacing 'promiscuous' single guide RNAs (sgRNAs) with modified sgRNAs.

TL;DR: The efficiency, accuracy and safety of the approach presented suggest that it has potential to be used for the correction of heritable mutations in human embryos by complementing preimplantation genetic diagnosis.

TL;DR: It is found that Cpf1 could tolerate single or double mismatches in the 3′ Pam-distal region, but not in the 5′ PAM-proximal region, and off-target effects were completely abrogated by using preassembled, recombinant CpF1 ribonucleoproteins.

TL;DR: CjCas9, delivered via AAV, induces targeted mutations at high frequencies in mouse muscle cells or retinal pigment epithelium (RPE) cells, and reduces the size of laser-induced choroidal neovascularization, suggesting that in vivo genome editing with Cj Cas9 is a new option for the treatment of age-related macular degeneration.