Engineered Mouse Sentinel System for Monitoring Animal Facilities

摘要 动物研究机构的主要使命是确保动物的健康和福祉， 人道利用动物研究结果的完整性、可靠性和可重复性。实现 为此，设施部署了各种方法，包括仔细控制动物协议，周到的工程， 设施，以及对传染性病原体的勤奋筛查。尽管尽了最大努力，这些方法不能 全面保护动物免受所有可能的脆弱性;因此，需要强大的监测工具 提醒护理人员注意NIH FOA PAR-19-266中强调的设施问题。拟议工作 将开发一种新的工具，通过嵌入一种基因， 报告系统。Cell出租公司发明了基于CRISPR-Cas9的新技术， 能够在哨兵小鼠的基因组DNA中进行生理事件的时间分辨记录。通过 通过在小鼠肠道中表达该系统，记录的事件通过快速且廉价的方法被有效地“读取”。 对从哨兵小鼠收集的粪便DNA进行测序。在两个目标中，该提案寻求1）建立一个基础系统 在小鼠中，可以与触发系统组合以制造哨兵，以及2）校准 使用强力霉素作为模型哨兵中的触发事件的基础系统。完成拟议的工作 第一阶段将为第二阶段监测生理压力的触发系统的工程设计提供基础 反应，炎症和暴露于病原体在新的哨兵小鼠。这一新工具将解决关键的 通过提供一种监测压力和健康状况的方法，满足动物设施未满足的需求， 老鼠，但否则不会被发现。

ABSTRACT The primary mission of animal research facilities is assuring the health and well-being of its animals for ensuring the integrity, reliability, and reproducibility of research results obtained by humanely using animals. To achieve this, facilities deploy a variety of methods, including careful control of animal protocols, thoughtful engineering of facilities, and diligent screening for infectious agents. Despite best efforts, these methods cannot comprehensively protect animals from all possible vulnerabilities; therefore, robust monitoring tools are needed to alert caretakers to problems in their facilities as highlighted in the NIH FOA PAR-19-266. The proposed work will develop a new tool that monitors the health and well being of mice in animal facilities by embedding a genetic reporting system into sentinel mice. Cellgorithmics, has invented new CRISPR-Cas9-based technologies that enable time-resolved recording of physiological events to be made in the genomic DNA of sentinel mice. By expressing the system in the mouse intestine, the recorded events are effectively “read” by rapid and inexpensive sequencing of fecal DNA collected from sentinel mice. In two Aims, the proposal seeks to 1) build a base system in mice that can be combined with triggering systems to make sentinels, and 2) calibrate the response of the base system using doxycycline as a triggering event in a model sentinel. Completion of the proposed work in Phase I will provide the basis for the engineering of triggering systems in Phase II to monitor physiological stress responses, inflammation, and exposure to pathogens in new sentinel mice. This new tool will address critical unmet needs of animal facilities by providing a way of monitoring stress and health conditions that directly affect mice, but would otherwise go undetected.