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Nat Biotechnol DOI:10.1038/s41587-019-0134-y

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

Publication TypeJournal Article
Year of Publication2019
AuthorsHuang, TP, Zhao, KT, Miller, SM, Gaudelli, NM, Oakes, BL, Fellmann, C, Savage, DF, Liu, DR
JournalNat Biotechnol
Date Published2019 May 20
ISSN1546-1696
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.

DOI10.1038/s41587-019-0134-y
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/31110355?dopt=Abstract

Alternate JournalNat. Biotechnol.
PubMed ID31110355
Grant ListU01 AI142756 / / Division of Intramural Research, National Institute of Allergy and Infectious Diseases (Division of Intramural Research of the NIAID) /
R01 EB022376 / / U.S. Department of Health & Human Services | NIH | National Institute of Biomedical Imaging and Bioengineering (NIBIB) /
R35 GM118062 / / U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS) /