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

• DOI: 10.1038/s41467-018-05262-w
• Publication Date: 2018-07-18T12:10:05Z
• Authors: Hiroki Sasaguri; Kenichi Nagata; Misaki Sekiguchi; Ryo Fujioka; Yukio Matsuba; Shoko Hashimoto; Kaori Sato; Deepika Kurup; Takanori Yokota; Takaomi C. Saido
• Keywords: Gene Editing; Male; Mice, Knockout; Mice, Inbred ICR; 0303 health sciences; Base Sequence; Science; Q; Reproducibility of Results; Article; Mice, Inbred C57BL; 03 medical and health sciences; Cytidine Deaminase; Sequence Homology, Nucleic Acid; Mutation; Presenilin-1; Animals; Female; CRISPR-Cas Systems; Base Pairing

AbstractBase 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.