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Circularly permuted and PAM-modified Cas9 variants broaden the targeting scope of base editors.

ABSTRACT: Base editing requires that the target sequence satisfy the protospacer adjacent motif requirement of the Cas9 domain and that the target nucleotide be located within the editing window of the base editor. To increase the targeting scope of base editors, we engineered six optimized adenine base editors (ABEmax variants) that use SpCas9 variants compatible with non-NGG protospacer adjacent motifs. To increase the range of target bases that can be modified within the protospacer, we use circularly permuted Cas9 variants to produce four cytosine and four adenine base editors with an editing window expanded from ~4-5 nucleotides to up to ~8-9 nucleotides and reduced byproduct formation. This set of base editors improves the targeting scope of cytosine and adenine base editing.

SUBMITTER: Huang TP

PROVIDER: S-EPMC6551276 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Circularly permuted and PAM-modified Cas9 variants broaden the targeting scope of base editors.

Huang Tony P TP Zhao Kevin T KT Miller Shannon M SM Gaudelli Nicole M NM Oakes Benjamin L BL Fellmann Christof C Savage David F DF Liu David R DR

Nature biotechnology 20190520 6

Base editing requires that the target sequence satisfy the protospacer adjacent motif requirement of the Cas9 domain and that the target nucleotide be located within the editing window of the base editor. To increase the targeting scope of base editors, we engineered six optimized adenine base editors (ABEmax variants) that use SpCas9 variants compatible with non-NGG protospacer adjacent motifs. To increase the range of target bases that can be modified within the protospacer, we use circularly ...[more]

PMID: 31110355

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