Ontology highlight
ABSTRACT:
SUBMITTER: Vakulskas CA
PROVIDER: S-EPMC6107069 | biostudies-literature | 2018 Aug
REPOSITORIES: biostudies-literature
Vakulskas Christopher A CA Dever Daniel P DP Rettig Garrett R GR Turk Rolf R Jacobi Ashley M AM Collingwood Michael A MA Bode Nicole M NM McNeill Matthew S MS Yan Shuqi S Camarena Joab J Lee Ciaran M CM Park So Hyun SH Wiebking Volker V Bak Rasmus O RO Gomez-Ospina Natalia N Pavel-Dinu Mara M Sun Wenchao W Bao Gang G Porteus Matthew H MH Behlke Mark A MA
Nature medicine 20180806 8
Translation of the CRISPR-Cas9 system to human therapeutics holds high promise. However, specificity remains a concern especially when modifying stem cell populations. We show that existing rationally engineered Cas9 high-fidelity variants have reduced on-target activity when using the therapeutically relevant ribonucleoprotein (RNP) delivery method. Therefore, we devised an unbiased bacterial screen to isolate variants that retain activity in the RNP format. Introduction of a single point mutat ...[more]