PT  - JOURNAL ARTICLE
AU  - Guodong Wang
AU  - Qi Li
AU  - Junjie Xu
AU  - Shuai Zhao
AU  - Rui Zhou
AU  - Zhenkang Chen
AU  - Wentong Jiang
AU  - Xue Gao
AU  - Shuang Zhou
AU  - Zhiyu Chen
AU  - Quanzhi Sun
AU  - Chengyuan Ma
AU  - Lin Chen
AU  - Bihan Shi
AU  - Ying Guo
AU  - Haiyan Wang
AU  - Xia Wang
AU  - Huaiye Li
AU  - Tao Cai
AU  - Yibing Wang
AU  - Zhineng Chen
AU  - Fengchao Wang
AU  - Qinghua Liu
TI  - Somatic Genetics Analysis of Sleep in Adult Mice
AID  - 10.1523/JNEUROSCI.0089-22.2022
DP  - 2022 Jul 13
TA  - The Journal of Neuroscience
PG  - 5617--5640
VI  - 42
IP  - 28
4099  - http://www.jneurosci.org/content/42/28/5617.short
4100  - http://www.jneurosci.org/content/42/28/5617.full
SO  - J. Neurosci.2022 Jul 13; 42
AB  - Classical forward and reverse mouse genetics require germline mutations and, thus, are unwieldy to study sleep functions of essential genes or redundant pathways. It is also time-consuming to conduct EEG/EMG-based mouse sleep screening because of labor-intensive surgeries and genetic crosses. Here, we describe a highly accurate SleepV (video) system and adeno-associated virus (AAV)-based adult brain chimeric (ABC)-expression/KO platform for somatic genetics analysis of sleep in adult male or female mice. A pilot ABC screen identifies CREB and CRTC1, of which constitutive or inducible expression significantly reduces quantity and/or quality of non-rapid eye movement sleep. Whereas ABC-KO of exon 13 of Sik3 by AAV-Cre injection in Sik3-E13flox/flox adult mice phenocopies Sleepy (Sik3Slp/+) mice, ABC-CRISPR of Slp/Sik3 reverses hypersomnia of Sleepy mice, indicating a direct role of SLP/SIK3 kinase in sleep regulation. Multiplex ABC-CRISPR of both orexin/hypocretin receptors causes narcolepsy episodes, enabling one-step analysis of redundant genes in adult mice. Therefore, this somatic genetics approach should facilitate high-throughput analysis of sleep regulatory genes, especially for essential or redundant genes, in adult mice by skipping mouse development and minimizing genetic crosses.SIGNIFICANCE STATEMENT The molecular mechanisms of mammalian sleep regulation remain unclear. Classical germline mouse genetics are unwieldy to study sleep functions of essential genes or redundant pathways. The EEG/EMG-based mouse sleep screening is time-consuming because of labor-intensive surgeries and lengthy genetic crosses. To overcome these “bottlenecks,” we developed a highly accurate video-based sleep analysis system and adeno-associated virus-mediated ABC-expression/KO platform for somatic genetics analysis of sleep in adult mice. These methodologies facilitate rapid identification of sleep regulatory genes, but also efficient mechanistic studies of the molecular pathways of sleep regulation in mice.