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， 由壁虱传播的伯氏疏螺旋体细菌引起。尽管人类和其他大型 哺乳动物可以感染伯氏杆菌，以便在野外完成其生命周期。 细菌依赖于啮齿动物的蓄水池，其中最主要的是白脚白脚拟青霉。 鹿鼠。白斑巴氏杆菌在莱姆病和其他几种壁虱传播疾病中的作用是 类似于蝙蝠作为SARS冠状病毒和埃博拉病毒的蓄水池。在本建议书中 我们继续发展白纹伊蚊作为一种新兴的遗传模型系统进行研究。 通过保持和扩大基因组和生物学上的 这一物种的资源。这些资源是任何以基因为中心的 Permyscus属的实验。这项提案的主要目标是确定 分离影响白假单胞菌作为细菌宿主能力的遗传因素 勃格多费里。水库能力的特征是通过感染和感染的流行率来衡量 从幼虫蜕皮的若虫队列中相应的细菌负荷 以前以实验感染的鹿鼠为食。次要终端包括 动物和选定宿主血液和皮肤中细菌的消长 反应，如对该制剂的抗体和组织炎症，在一段时间内 感染。通常情况下，进行大规模基因分型和/或 一种新出现的啮齿动物模型中的基因杂交。在这里，我们展示了我们对P. Leucopus与来自长期封闭群体的低通短读序列 鹿鼠可以被用来准确地将SNP和单倍型基因归因于基因组- 规模很大。然后，这些基因类型被用来识别与显著相关的基因 白纹伊蚊作为伯氏杆菌和其他疾病主要宿主的能力 探员们。最后，将通过CRISPR/Cas9基因敲除来验证已识别基因的子集 由这一属的转基因先驱者带头研究的白纹平革菌。这个 识别储层能力中介基因可能会提出更好的干预措施 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.