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

Hiroki Sasaguri (), Kenichi Nagata, Misaki Sekiguchi, Ryo Fujioka, Yukio Matsuba, Shoko Hashimoto, Kaori Sato, Deepika Kurup, Takanori Yokota and Takaomi C. Saido ()
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Hiroki Sasaguri: RIKEN Center for Brain Science
Kenichi Nagata: RIKEN Center for Brain Science
Misaki Sekiguchi: RIKEN Center for Brain Science
Ryo Fujioka: RIKEN Center for Brain Science
Yukio Matsuba: RIKEN Center for Brain Science
Shoko Hashimoto: RIKEN Center for Brain Science
Kaori Sato: RIKEN Center for Brain Science
Deepika Kurup: RIKEN Center for Brain Science
Takanori Yokota: Tokyo Medical and Dental University
Takaomi C. Saido: RIKEN Center for Brain Science

Nature Communications, 2018, vol. 9, issue 1, 1-8

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-05262-w

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