Highly efficient base editing in Staphylococcus aureus using an engineered CRISPR RNA-guided cytidine deaminase. Issue 12 (2nd March 2018)

• Record Type: Journal Article
• Authors: Gu, Tongnian; Zhao, Siqi; Pi, Yishuang; Chen, Weizhong; Chen, Chuanyuan; Liu, Qian; Li, Min; Han, Dali; Ji, Quanjiang
• Is Part Of: Chemical science. Volume 9:Issue 12(2018)
• Page Start: 3248
• Page End: 3253
• Publication Date: 2018-03-02
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c8sc00637g
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : The base editor pnCasSA-BEC enables highly efficient base editing in Staphylococcus aureus . Abstract : Novel therapeutic means against Staphylococcus aureus infections are urgently needed due to the emergence of drug-resistant S. aureus . We report the development of a CRISPR RNA-guided cytidine deaminase (pnCasSA–BEC), enabling highly efficient gene inactivation and point mutations in S. aureus . We engineered a fusion of a Cas9 nickase (Cas9D10A) and a cytidine deaminase (APOBEC1) that can be guided to a target genomic locus for gene inactivation via generating a premature stop codon. The pnCasSA–BEC system nicks the non-edited strand of the genomic DNA, directly catalyzes the conversion of cytidine (C) to uridine (U), and relies on DNA replication to achieve C → T (G → A) conversion without using donor repair templates. The development of the base-editing system will dramatically accelerate drug-target exploration in S. aureus and provides critical insights into the development of base-editing tools in other microbes.