Sequencing Mutant Mice With Altered Cocaine Responses

 描述(由申请人提供)：这项提案的目标是发现调节对成瘾药物可卡因敏感性的新基因变异。发现调节成瘾的新基因和等位基因将有助于发现成瘾的机制成分 可作为治疗和干预目标的途径。我们采取了一种先进的基因突变方法，以发现调节对可卡因、多动和焦虑的急性反应的变异，所有这些都是成瘾行为的预测因素。在模型动物中进行基于突变的筛选是完全没有偏见的，50多年来一直是生物学的基础。我们进行了迄今为止最大的可卡因反应和公开领域行为筛查之一。从这个超过17,000只小鼠的筛查中，我们收集了10个突变，并将其中3个突变定位到大的染色体间隔。现在，我们建议利用下一代测序技术来找到导致这些突变体表型差异的确切突变。我们还将携带我们的RNA-Seq，以了解在这些突变体中被干扰的转录程序。我们将利用外显子组、全基因组和RNA测序来研究这些突变体，进行综合的多步骤下一代测序方法。该项目的成功完成将导致检测成瘾易感性的可行因果变异，并识别受该变异干扰的途径。这个项目在发现新的基因、等位基因和途径方面具有巨大的潜力。 来调节可卡因的反应。从这些信息中，我们将能够识别新的成瘾神经底物，发现新的分子成分，并建立更好的机制理解可卡因的反应及其与潜在可吸毒靶点的接近程度。我们还将为成瘾研究界提供改变药物反应的新小鼠模型。

 DESCRIPTION (provided by applicant): The goal of this proposal is to discover novel genetic variants that regulate sensitivity to an addictive drug, cocaine. Discovery of novel genes and alleles that mediate addiction will lend itself to discovery of mechanistic components in addiction pathways that can be targeted for treatment and intervention. We have taken a forward genetic mutagenesis approach to discover variants that regulate acute response to cocaine, hyperactivity and anxiety, all predictors of addictive behavior. Mutagenesis based screening in model animals is completely unbiased and has been bedrock in biology for over 50 years. We have conducted one of the largest screens to date for cocaine response and open field behaviors. From this screen of over 17,000 mice we have a collection of ten mutants and have genetically mapped three of these mutants to large chromosomal intervals. Now, we propose to leverage next-generation sequencing technology to find the exact mutation responsible for the phenovariance in these mutants. We will also carry our RNA-Seq to understand the transcriptomic program that is perturbed in these mutants. We will carry out an integrated multistep next- generation sequencing approach utilizing exome, whole genome, and RNA sequencing to study these mutants. Successful completion of this project will lead to detection of viable causal variants for addiction liability and identification of the pathway perturbed by th variant. This project has significant potential for discovery of novel genes, alleles, and pathways that regulate cocaine responses. From this information we will be able to identify new neural substrates of addiction, discover novel molecular components and establish a better mechanistic understanding of cocaine response and its proximity to potential druggable targets. We will also provide the addiction research community with new mouse models for altered drug response.