Mammalian reverse genetics without crossing reveals Nr3a as a short-sleeper gene
Sunagawa, Genshiro A., Sumiyama, Kenta, Ukai-Tadenuma, Maki, Perrin, Dimitri, Fujishima, Hiroshi, Ukai, Hideki, Nishimura, Osamu, Shi, Shoi, Ohno, Rei-ichiro, Narumi, Ryohei, Shimizu, Yoshihiro, Tone, Daisuke, Ode, Koji L., Kuraku, Shigehiro, & Ueda, Hiroki R. (2016) Mammalian reverse genetics without crossing reveals Nr3a as a short-sleeper gene. Cell Reports, 14, pp. 1-16.
The identification of molecular networks at the system level in mammals is accelerated by next-generation mammalian genetics without crossing, which requires both the efficient production of whole-body biallelic knockout (KO) mice in a single generation and high-performance phenotype analyses. Here, we show that the triple targeting of a single gene using the CRISPR/Cas9 system achieves almost perfect KO efficiency (96%–100%). In addition, we developed a respiration-based fully automated noninvasive sleep phenotyping system, the Snappy Sleep Stager (SSS), for high-performance (95.3% accuracy) sleep/wake staging. Using the triple-target CRISPR and SSS in tandem, we reliably obtained sleep/wake phenotypes, even in double-KO mice. By using this system to comprehensively analyze all of the N-methyl-D-aspartate (NMDA) receptor family members, we found Nr3a as a short-sleeper gene, which is verified by an independent set of triple-target CRISPR. These results demonstrate the application of mammalian reverse genetics without crossing to organism-level systems biology in sleep research.
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|Item Type:||Journal Article|
|Divisions:||Current > Schools > School of Electrical Engineering & Computer Science
Current > QUT Faculties and Divisions > Science & Engineering Faculty
|Copyright Owner:||Copyright 2016 The Authors|
|Copyright Statement:||This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).|
|Deposited On:||10 Jan 2016 23:33|
|Last Modified:||11 Jan 2016 23:19|
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