Efficient generation of mouse models of human diseases via ABE- and BE-mediated base editing

Nat Commun. 2018 Jun 14;9(1):2338. doi: 10.1038/s41467-018-04768-7.

Abstract

A recently developed adenine base editor (ABE) efficiently converts A to G and is potentially useful for clinical applications. However, its precision and efficiency in vivo remains to be addressed. Here we achieve A-to-G conversion in vivo at frequencies up to 100% by microinjection of ABE mRNA together with sgRNAs. We then generate mouse models harboring clinically relevant mutations at Ar and Hoxd13, which recapitulates respective clinical defects. Furthermore, we achieve both C-to-T and A-to-G base editing by using a combination of ABE and SaBE3, thus creating mouse model harboring multiple mutations. We also demonstrate the specificity of ABE by deep sequencing and whole-genome sequencing (WGS). Taken together, ABE is highly efficient and precise in vivo, making it feasible to model and potentially cure relevant genetic diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / chemistry
  • Animals
  • Animals, Newborn
  • Cell Line, Tumor
  • Disease Models, Animal*
  • Female
  • Genome, Human
  • Genotype
  • Green Fluorescent Proteins / chemistry
  • High-Throughput Nucleotide Sequencing
  • Homeodomain Proteins / genetics*
  • Humans
  • Male
  • Mice
  • Mutation
  • Protein Domains
  • RNA, Guide, CRISPR-Cas Systems
  • RNA, Messenger / metabolism
  • Receptors, Androgen / genetics*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Transcription Factors / genetics*

Substances

  • AR protein, mouse
  • Homeodomain Proteins
  • Hoxd13 protein, mouse
  • RNA, Messenger
  • Receptors, Androgen
  • Transcription Factors
  • Green Fluorescent Proteins
  • Adenine