Introduction of pathogenic mutations into the mouse Psen1 gene by Base Editor and Target-AID

Nat Commun. 2018 Jul 24;9(1):2892. doi: 10.1038/s41467-018-05262-w.

Abstract

Base Editor (BE) and Target-AID (activation-induced cytidine deaminase) are engineered genome-editing proteins composed of Cas9 and cytidine deaminases. These base-editing tools convert C:G base pairs to T:A at target sites. Here, we inject either BE or Target-AID mRNA together with identical single-guide RNAs (sgRNAs) into mouse zygotes, and compare the base-editing efficiencies of the two distinct tools in vivo. BE consistently show higher base-editing efficiency (10.0-62.8%) compared to that of Target-AID (3.4-29.8%). However, unexpected base substitutions and insertion/deletion formations are also more frequently observed in BE-injected mice or zygotes. We are able to generate multiple mouse lines harboring point mutations in the mouse presenilin 1 (Psen1) gene by injection of BE or Target-AID. These results demonstrate that BE and Target-AID are highly useful tools to generate mice harboring pathogenic point mutations and to analyze the functional consequences of the mutations in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Pairing / genetics*
  • Base Sequence
  • CRISPR-Cas Systems*
  • Cytidine Deaminase / genetics*
  • Cytidine Deaminase / metabolism
  • Female
  • Gene Editing / methods*
  • Male
  • Mice, Inbred C57BL
  • Mice, Inbred ICR
  • Mice, Knockout
  • Mutation*
  • Presenilin-1 / genetics*
  • Presenilin-1 / metabolism
  • Reproducibility of Results
  • Sequence Homology, Nucleic Acid

Substances

  • Presenilin-1
  • presenilin 1, mouse
  • Cytidine Deaminase