A single CRISPR base editor to induce simultaneous C-to-T and A-to-G mutations

Rina C. Sakata, Soh Ishiguro, Hideto Mori, Mamoru Tanaka, Motoaki Seki, Nanami Masuyama, Keiji Nishida, Hiroshi Nishimasu, Akihiko Kondo, Osamu Nureki, Masaru Tomita, Hiroyuki Aburatani, Nozomu Yachie

Research output: Contribution to journalArticlepeer-review

Abstract

While several Cas9-derived base editors have been developed to induce either C-to-T or A-to-G mutation at target genomic sites, the possible genome editing space when using the current base editors remains limited. Here, we present a novel base editor, Target-ACE, which integrates the abilities of both of the previously developed C-to-T and A-to-G base editors by fusing an activation-induced cytidine deaminase (AID) and an engineered tRNA adenosine deaminase (TadA) to a catalytically impaired Streptococcus pyogenes Cas9. In mammalian cells, Target-ACE enabled heterologous editing of multiple bases in a small sequence window of target sites with increased efficiency compared with a mixture of two relevant base editor enzymes, each of which may block the same target DNA molecule from the other. Furthermore, by modeling editing patterns using deep sequencing data, the editing spectra of Target-ACE and other base editors were simulated across the human genome, demonstrating the highest potency of Target-ACE to edit amino acid coding patterns. Taking these findings together, Target-ACE is a new tool that broadens the capabilities for base editing for various applications.

Original languageEnglish
JournalUnknown Journal
DOIs
Publication statusPublished - 2019 Aug 8

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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