Enhanced genome editing efficiency of CRISPR PLUS: Cas9 chimeric fusion proteins
Jongjin Park, Ji-Young Yoon, Daekee Kwon, Mi-Jung Han, Sunmee Choi, Slki Park, Junghyuk Lee, Ki-Wook Lee, John J. Lee, Seunghee Lee, Kyung‐Sun Kang, Sunghwa Choe
Abstract
Efforts to improve CRISPR-Cas9 genome editing systems for lower off-target effects are mostly at the cost of its robust on-target efficiency. To enhance both accuracy and efficiency, we created chimeric SpyCas9 proteins fused with the 5'-to-3' exonuclease Recombination J (RecJ) or with GFP and demonstrated that transfection of the pre-assembled ribonucleoprotein of the two chimeric proteins into human or plant cells resulted in greater targeted mutagenesis efficiency up to 600% without noticeable increase in off-target effects. Improved activity of the two fusion proteins should enable editing of the previously hard-to-edit genes and thus readily obtaining the cells with designer traits.