Genetic architecture of host response to tickborne disease in Peromyscus leucopus

项目摘要 莱姆病是北美最常见的传染病之一， 由蜱传伯氏疏螺旋体引起虽然人类和其他大型 哺乳动物可以被B感染。burgdorferi为了在野外完成它的生命周期， 细菌依赖于啮齿动物的水库，主要的一个是白足鼠，白足鼠 鹿鼠白足拟杆菌在莱姆病和其他几种蜱传疾病中的作用是 类似于蝙蝠作为SARS冠状病毒和埃博拉病毒的宿主。本提案中 我们继续发展P. leucopus作为研究的新兴遗传模型系统 传染病和其他疾病的基因组和生物 这个物种的资源。这些资源是任何以基因为重点的 实验在Peromyscus属。该提案的主要目标是确定 分离影响白柄侧耳作为B库的能力的遗传因素。 burgdorferi。储库能力的特征被测量为感染的患病率， 在一群从幼虫蜕皮的蝗虫中， 之前以实验感染的鹿鼠为食次要终点包括 动物和选定宿主血液和皮肤中细菌的生长和衰退 反应，如抗体的代理和炎症的组织，随着时间的推移， 感染进行大规模的基因分型和/或 在一个新兴的啮齿动物模型中进行基因杂交。在这里，我们表明，我们的基因组组装P。 leucopus与来自长期封闭菌落的低通短读段序列一致 鹿鼠可以用来准确地估算基因组上的SNP和单倍型基因型- 大规模然后，这些基因型被用来识别有助于显著的 白足拟青霉是B的主要贮存宿主。Burgdorferi和其他疾病 剂.最后，将通过CRISPR/Cas9基因敲除来验证已鉴定基因的子集。 在P. leucopus中，由该属转基因技术的先驱者带头。的 储层能力介导基因的鉴定可能提示更好的干预措施， 阻断传播并为人类感染的管理提供见解。

Project Summary Lyme disease, one of the most commonly reported infectious diseases in North America, is caused by the tick-borne bacterium Borreliella burgdorferi. Although humans and other large mammals can be infected by B. burgdorferi, in order to complete its life-cycle in the wild the bacteria relies on rodent reservoirs, the major one being Peromyscus leucopus, the white-footed deermouse. The role of P. leucopus in Lyme disease and several other tick-borne diseases is analogous to that of bats as reservoirs for SARS coronaviruses and Ebola virus. In this proposal we continue the development of P. leucopus as an emerging genetic model system for the study of infectious and other diseases by maintaining and expanding genomic and biological resources for this species. These resources are the starting point for any gene-focused experiments in the Peromyscus genus. The primary goal of this proposal is to identify segregating genetic factors that impact the competence of P. leucopus as a reservoir of B. burgdorferi. The trait of reservoir competence is measured as the prevalence of infection and corresponding bacterial burdens among a cohort of nymphs that had molted from larvae previously fed on experimentally-infected deermice. Secondary endpoints include rates of growth and decline of the bacteria in the blood and skin of the animals and selected host responses, such as antibodies to the agent and inflammation of tissues, over the time course of the infection. It would normally be extremely difficult to carry-out large-scale genotyping and/or genetic crosses in an emerging rodent model. Here we show that our genome assembly for P. leucopus in concert with low pass short read sequences from a long-term closed colony of deermice can be leveraged to accurately impute SNP and haplotype genotypes on a genome- wide scale. These genotypes are then used to identify genes contributing to the remarkable capacity of P. leucopus to serve as a key reservoir host for B. burgdorferi and other disease agents. Finally, a subset of identified genes will be validated via CRISPR/Cas9 gene knock-outs in P. leucopus spearheaded by the person who pioneered transgenics for this genus. The identification of reservoir competence mediating genes may suggest better interventions to block transmission and provide insights into the management of human infections.