Enhanced genome editing efficiency of CRISPR PLUS: Cas9 chimeric fusion proteins

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...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Jongjin Park, Jiyoung Yoon, Daekee Kwon, Mi-Jung Han, Sunmee Choi, Slki Park, Junghyuk Lee, Kiwook Lee, Jaehwan Lee, Seunghee Lee, Kyung-Sun Kang, Sunghwa Choe
Formato: article
Lenguaje:EN
Publicado: Nature Portfolio 2021
Materias:
R
Q
Acceso en línea:https://doaj.org/article/b70942f95a9247b8bb22b7e0f6eacb48
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
Descripción
Sumario: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.