开发和特征小鼠适应的复合SARS-CoV-2表达记者基因的开发和特征
bioRxiv : the preprint server for biology
|February 27, 2026
概括
我们开发了一种重组的SARS-CoV-2 (严重急性呼吸系统综合征冠状病毒2),具有用于跟踪小鼠感染的记者基因. 这种新工具允许对病毒传播进行非侵入性监测,并评估野生类型小鼠的治疗方法.
科学领域:
- 病毒学 病毒学
- 传染性疾病 传染性疾病
- 生物技术是生物技术.
背景情况:
- 转基因K18-hACE2小鼠是SARS-CoV-2研究的标准,但具有局限性.
- 适应于老鼠的SARS-CoV-2 (MA30) 允许野生型老鼠感染,但缺乏易于跟踪的方法.
- 现有的模型需要二次方法来监测病毒感染,使研究复杂化.
研究的目的:
- 开发一种重组的SARS-CoV-2 (rSARS-CoV-2) 菌株,以有效跟踪病毒感染.
- 为了使SARS-CoV-2感染动态在体外,体外和体内进行非侵入性监测.
- 为了促进预防性疫苗和治疗策略在野生类型小鼠的评估.
主要方法:
- 开发了一种复合MA30SARS-CoV-2菌株,表达光 (mCherry) 和/或光酶 (nanoluciferase,Nluc) 记者基因.
- 在试验室使用A549 hACE2和Vero AT细胞评估病毒衰减和复制动力学.
- 通过生物发光成像在WT C57BL/6和BALB/c小鼠中评估了体内感染动态,体重减轻和病毒载荷.
主要成果:
- 记者基因插入在体外引起了轻微的病毒衰减 (~0.5-1.0-log较低的标位),但维持了复制动力学.
- 在体内,报告者表达的rSARS-CoV-2 MA30诱导了暂时的体重减轻,与致命的WT MA30感染不同.
- 生物发光成像显示,感染后2天肺复制达到峰值,在4天内消失,与组织病毒负载相关.
结论:
- 具有记者基因的重组SARS-CoV-2 MA30可有效地在野生类型小鼠中进行体外,体外和体内感染跟踪.
- 该系统绕过了WT MA30所需的二次监控方法的需求.
- 报告员表达的rSARS-CoV-2 MA30适合在各种小鼠模型中进行非侵入性监测,评估疫苗和治疗方法.
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