Engineered Mouse Sentinel System for Monitoring Animal Facilities

用于监测动物设施的工程小鼠哨兵系统

• 批准号: 10323482
• 负责人: Ryan Clarke
• 金额: $ 23.7万
• 依托单位: CELLGORTHMICS, INC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323482
• 关键词: Address; Affect; Animals; Biological; Biomedical Research; CRISPR/Cas technology; Cells; Communicable Diseases; Control Animal; Corticosterone; DNA; DNA Sequence; DNA sequencing; Detection; Development; Development Plans; Doxycycline; Engineering; Ensure; Environment; Epithelial Cells; Event; Exposure to; Feces; Future; Gene Expression; Genes; Genetic; Genetic Engineering; Genome; Genomic DNA; Health; Infection; Infectious Agent; Inflammation; Intellectual Property; Interferons; Intestines; Laboratory Animal Production and Facilities; Link; Methods; Mission; Modeling; Monitor; Mus; NR5A2 gene; Nature; Pain; Personal Satisfaction; Phase; Physiologic Monitoring; Physiological; Physiology; Preparation; Protocols documentation; Reagent; Records; Regimen; Reporting; Reproducibility; Research; Resources; Sampling; Sentinel; Series; Signal Pathway; Signal Transduction; Standardization; Stimulus; Stress; System; Technology; Testing; Ticks; Time; Transgenes; United States National Institutes of Health; Work; animal facility; base; biological adaptation to stress; cost; environmental stressor; experience; extracellular; interleukin-22; intestinal epithelium; mouse genome; new technology; novel; pathogen; pathogenic bacteria; promoter; response; screening; tool; transcription factor

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.

抽象的 动物研究设施的主要任务是确保其动物的健康和福祉 人文使用动物获得的研究结果的完整性，可靠性和可重复性。实现 这是部署各种方法的设施，包括仔细控制动物协议，周到的工程 设施，以及对传染剂的勤奋筛查。尽管尽了最大的努力，这些方法不能 全面保护动物免受所有可能的脆弱性；因此，需要强大的监视工具 正如NIH FOA PAR-19-266中所强调的那样，要提醒看护者的设施问题。拟议的工作 将开发一种新的工具，通过嵌入通用物质来监测动物设施中的老鼠的健康和健康 向哨兵小鼠报告系统。 Cellgorithmics，已经发明了新的基于CRISPR-CAS9的技术 可以在哨兵小鼠的基因组DNA中对物理事件进行时间分辨记录。经过 在鼠标肠中表达系统，记录的事件通过快速且廉价的有效地“读取” 从哨兵小鼠收集的粪便DNA测序。在两个目标中，该提案寻求1）建立一个基本系统 在可以与触发系统结合的小鼠中，2）校准 基本系统使用强力霉素作为模型前哨中的触发事件。完成拟议的工作 第一阶段将为II阶段触发系统的工程提供基础，以监测身体压力 新哨兵小鼠的反应，炎症和暴露于病原体。这个新工具将解决关键 通过提供一种直接影响压力和健康状况的方式，未满足动物设施的需求 老鼠，但否则会不会被发现。