构建时空可调控的人类遗传疾病模型

o overcome the limitation that most point mutations introduced in previous studies to mimic human genetic diseases were expressed systemically, restricting investigations with temporal and spatial specificity, this study utilized CRISPR/Cas9 and LoxP/Cre systems to develop a spatiotemporally controllable PER2S659G mouse model. Using genome editing, a PER2S659Gflox/+ mouse with a conditionally expressed mutation was generated. To validate the model\'s construction, this mouse was first crossed with CMV-Cre mice to establish a systemic PER2S659G expression model. The efficacy of the model was assessed via genotyping, sequencing validation, and behavioral experiments, including wheel-running analysis. Results confirmed the successful establishment of the model. Wheel-running analysis revealed that systemic PER2S659G expression significantly shortened the circadian period to 22.04 ± 0.22 hours, approximately 1.5 hours shorter than the control groups (Cmv-Cre: 23.85 ± 0.06 hours; PER2S659Gflox/+: 23.64 ± 0.16 hours). Additionally, systemic expression advanced activity onset by approximately 3.5 hours (3.77 ± 0.6 hours) compared to the control groups (Cmv-Cre: -0.13 ± 0.20 hours; PER2S659Gflox/+: 0.13 ± 0.17 hours). This study successfully established a spatiotemporally controllable PER2S659G mouse model, offering a versatile tool for investigating the spatiotemporal-specific effects of circadian clock gene mutations through mating with different CRE lines and exploring their impact on sleep and activity rhythms.