CRISPR screens of population relevant genes governing toxicant resilience

人们对接触化学毒物的反应和影响有很大差异， 生物毒素因此，与不同个体暴露相关的不良后果风险 而且种群也有很大的差异基因与环境（G x E）的相互作用可能是一个重要的基础。 这些风险的差异。然而，我们仍然在很大程度上忽视了关键的遗传因素， 与特定毒素/毒物的机械关联。因此，我们降低风险的能力， 查明易受感染的个人和人群以便能够进行有效的预防努力， 有限公司目前鉴定G x E相互作用的方法依赖于遗传关联研究， 缺乏足够的力量来确定显着的关联，由于大量的遗传变异和小 暴露人群。我们建议，在一个根本不同的方法，首先系统地 确定影响对特定毒物/毒素的功能反应的常见人类变异体， 描述关键候选G X E相互作用，以便在相关个体和人群中进行有针对性的考虑。 我们将专注于1490个基因的功能询问，ToxVar集，其中包含聚合频率 迄今为止评估的所有人群中>0.1%的功能丧失突变， 与一种或多种有毒物质/毒素相互作用。我们认为，这些通常功能受损的 基因最有可能影响大部分人对毒素/毒物的反应。我们 将同时查询这1490个基因中每一个的功能中断对细胞的影响， 使用偶联的CRISPR筛选和单细胞毒理学相关基因对毒物的反应 表达靶点（scTRG）。我们将评估ToxVar-scTRGET方法，以在功能上识别 参与细胞对高人类毒性物质反应的相关和常见变异基因 在越来越多的生理相关的细胞模型的相关性。

People vary considerably in their response to and in the effects of exposure to chemical toxicants and biological toxins. As a result, the risk of adverse outcomes associated with exposure for different individuals and populations can be widely divergent. Gene by environment (G x E) interactions likely underlie a significant component of these risk differences. However, we remain largely ignorant of both the key genetic factors and the mechanistic association with specific toxins/toxicants. As a result, our capability to mitigate risk by the identification of susceptible individuals and populations to enable effective preventive efforts remain sorely limited. Current approaches to identify G x E interactions rely on genetic association studies which generally lack sufficient power to identify significant associations, due to the large number of genetic variants and small populations of exposed individuals. We propose, in a fundamentally different approach, to first systematically identify the common human variants which impact the functional response to a specific toxicant/toxin to delineate key candidate G X E interactions for targeted consideration in relevant individuals and populations. We will focus on functional interrogation of 1490 genes, the ToxVar set, which contain an aggregate frequency of loss of function mutations of >0.1% in all human populations assessed to date and previously identified as interacting with one or more toxicant/toxins. We contend that these commonly functionally compromised genes are most likely to impact human response to a toxin/toxicant in a significant proportion of people. We will simultaneously query the impact of functional disruption in each of these 1490 genes on the cellular response to a toxicant using coupled CRISPR screening and single cell toxicologically relevant gene expression targets (scTRGETs). We will evaluate the ToxVar-scTRGET approach to identify functionally relevant and commonly variant genes involved in cellular response to selected toxicants of high human relevance in increasingly physiologically relevant cell models.