Supplement: CRISPR screens of population relevant genes governing toxicant resilience

补充：CRISPR 筛选控制毒物抵抗力的群体相关基因

• 批准号: 10720972
• 负责人: CHRISTOPHER D VULPE
• 金额: $ 36.64万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-14 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10720972
• 关键词: Afferent Neurons; Alleles; Ataxia; CRISPR library; CRISPR screen; Candidate Disease Gene; Cardiac Myocytes; Categories; Cell Line; Cell model; Cells; Characteristics; Chromosome Mapping; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Dedications; Disabled Persons; Disease; Environment; Environmental Exposure; Environmental Impact; Environmental Pollutants; Environmental Risk Factor; Exposure to; Gene Expression; Genes; Genetic; Genetic Diseases; Genetic Transcription; Goals; Health; Hereditary Disease; Heterogeneity; Human; Individual; Introns; Laboratories; Length; Libraries; Mediating; Mendelian disorder; Mentorship; Motor; Mutation; Neurodegenerative Disorders; Phenotype; Population; Recording of previous events; Research; Research Assistant; Resources; Role; Spinal Ganglia; Stress; Students; System; Training; Trinucleotide Repeats; Undifferentiated; Variant; Work; autosome; candidate identification; career; career development; cell type; differential expression; disability; disabled students; experimental study; frataxin; gene correction; gene environment interaction; gene product; genetic variant; genome wide association study; graduate student; induced pluripotent stem cell; insight; loss of function; mutant; novel; parent grant; resilience; response; screening; single-cell RNA sequencing; stressor; toxicant; transcriptome sequencing; vector

Summary The goal of this supplement is to enhance diversity in the research workforce through support of a disabled graduate student in response to PA-21-071(Research Supplement to Promote Diversity in Health-Related Research). This supplement will enable the disabled student to work in the laboratory of the PI of parent grant by providing support for his stipend and tuition, as well as allowing the hiring of a dedicated technical research assistant for the components of the proposed work requiring fine motor capabilities. In addition, mentorship, career development support, targeted training efforts, and reasonable accommodations will enable the student to continue to pursue a scientific career while navigating the challenges presented by his disability. The parent grant focuses on GXE interactions with exogenous stressors but posited application to GXE interactions with endogenous stressors, such as a pre-existing monogenic genetic disorder, which is the focus of the supplement. We selected the monogenic disorder, Friedrich’s ataxia (FRDA), a progressive neurodegenerative disorder most commonly due to triplet repeat mutations in the frataxin gene. While triplet repeat length plays a role in the disease presentation, other unknown genetic and environmental factors clearly contribute. We will assess the capability of our novel targeted functional screening approach to identify genes which when disrupted could modulate the cellular phenotype of FRDA. As with the parent grant, the work will focus on the set of ~1490 genes with common LOF mutations in the population and will assess functional effect by assessing changes in the transcriptional phenotype of relevant cells. This work if successful could provide important insight into common LOF variants that impact the presentation and progression of Friedrich’s ataxia. This functional screening approach using transcriptional phenotypic endpoints could have general applicability to any genetic disease and enable identification of potentially functionally significant and population relevant genetic variants impacting the phenotypic manifestations of the disease.

概括 该增刊的目标是通过支持残疾人来增强研究队伍的多样性 研究生回应 PA-21-071（促进健康相关多样性的研究补充 研究）。该补助金将使残疾学生能够在家长资助的 PI 实验室工作 通过为他的津贴和学费提供支持，以及允许雇用专门的技术研究人员 助理负责拟议工作中需要精细运动能力的部分。此外，辅导、 职业发展支持、有针对性的培训工作和合理的住宿将使学生能够 继续追求科学事业，同时应对残疾带来的挑战。家长 拨款重点关注 GXE 与外源压力源的相互作用，但提出了 GXE 与 内源性压力源，例如预先存在的单基因遗传性疾病，这是补充剂的重点。 我们选择了单基因疾病弗里德里希共济失调 (FRDA)，这是一种最常见的进行性神经退行性疾病。 通常是由于 frataxin 基因中的三联体重复突变所致。虽然三联体重复长度在 疾病表现、其他未知的遗传和环境因素显然有影响。我们将评估 我们新颖的靶向功能筛选方法能够识别基因，这些基因在被破坏时可以 调节 FRDA 的细胞表型。与家长资助一样，这项工作将集中在 ~1490 人群中具有常见 LOF 突变的基因，并将通过评估基因的变化来评估功能效果 相关细胞的转录表型。这项工作如果成功的话可以提供重要的见解 影响弗里德里希共济失调表现和进展的常见 LOF 变异。这个功能性的 使用转录表型终点的筛选方法可以普遍适用于任何遗传 疾病并能够识别具有潜在功能意义和群体相关的遗传变异 影响疾病的表型表现。