期刊
CELL REPORTS
卷 14, 期 3, 页码 662-677出版社
CELL PRESS
DOI: 10.1016/j.celrep.2015.12.052
关键词
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类别
资金
- Innovative Cell Biology by Innovative Technology Program from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- MEXT/Japan Society for the Promotion of Science (JSPS) [25221004, 23115006]
- strategic programs for R&D (President's discretionary fund) of RIKEN
- RIKEN Center for Developmental Biology and RIKEN Quantitative Biology Center
- AMED Core Research for Evolutionary Medical Science and Technology (AMED-CREST), Japan AMED
- Core Research for Evolutionary Science and Technology (CREST)
- Japan Science and Technology Agency (JST)
- Program for the Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) from the MEXT
- Basic Science and Platform Technology Program for Innovative Biological Medicine from the Japan Agency for Medical Research and Development
- RIKEN Foreign Postdoctoral Researcher Program
- [26870858]
- [26113720]
- Grants-in-Aid for Scientific Research [26870858, 26640134, 14J06417, 26710014, 26113720] Funding Source: KAKEN
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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