Orthogonal Cas9-Cas9 chimeras provide a versatile platform for genome editing

Orthogonal Cas9-Cas9 chimeras provide a versatile platform for genome editing

The development of robust, versatile and accurate toolsets is critical to facilitate therapeutic genome editing applications. Here we establish RNA-programmable Cas9-Cas9 chimeras, in single- and dual-nuclease formats, as versatile genome engineering systems. In both of these formats, Cas9-Cas9 fusi...

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Bibliographic Details
Published in:Nature communications 2018-11, Vol.9 (1), p.4856-12, Article 4856
Main Authors: Bolukbasi, Mehmet Fatih, Liu, Pengpeng, Luk, Kevin, Kwok, Samantha F, Gupta, Ankit, Amrani, Nadia, Sontheimer, Erik J, Zhu, Lihua Julie, Wolfe, Scot A
Format: Article
Language:eng
Subjects:
Chimeras
Editing
Genomes
Nuclease
Nucleotide sequence
Ribonucleic acid
RNA
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Summary:The development of robust, versatile and accurate toolsets is critical to facilitate therapeutic genome editing applications. Here we establish RNA-programmable Cas9-Cas9 chimeras, in single- and dual-nuclease formats, as versatile genome engineering systems. In both of these formats, Cas9-Cas9 fusions display an expanded targeting repertoire and achieve highly specific genome editing. Dual-nuclease Cas9-Cas9 chimeras have distinct advantages over monomeric Cas9s including higher target site activity and the generation of predictable precise deletion products between their target sites. At a therapeutically relevant site within the BCL11A erythroid enhancer, Cas9-Cas9 nucleases produced precise deletions that comprised up to 97% of all sequence alterations. Thus Cas9-Cas9 chimeras represent an important tool that could be particularly valuable for therapeutic genome editing applications where a precise cleavage position and defined sequence end products are desirable.
ISSN:2041-1723
2041-1723