Noncoding DNA regulatory elements and Anopheles vector biology

非编码 DNA 调控元件和按蚊载体生物学

• 批准号: 10328247
• 负责人: Michelle M Riehle
• 金额: $ 34.75万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10328247
• 关键词: Africa; Alleles; Animals; Anopheles Genus; Anopheles gambiae; Behavior; Biological; Biological Assay; Biology; CRISPR/Cas technology; Cell Line; Chromosome Arm; Chromosomes; Code; Complex; Culicidae; DNA; Data; Detection; Dissection; Elements; Enhancers; Frequencies; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Enhancer Element; Genetic Markers; Genetic Polymorphism; Genetic Recombination; Genetic Variation; Genetic Vectors; Genome; Genomics; Genotype; Goals; Individual; Infection; Insecticide Resistance; Lead; Link; Malaria; Maps; Messenger RNA; MicroRNAs; Nature; Nucleotides; Organism; Outcome; Phenotype; Plant Roots; Plasmodium; Plasmodium falciparum; Population; Positioning Attribute; Predisposition; Prevalence; Proteins; Publishing; Regulatory Element; Resistance; Resolution; Resources; Science; Source; Susceptibility Gene; Technology; Testing; Time; Untranslated RNA; Variant; Vector Ecology; base; detection assay; disease phenotype; feeding; genetic analysis; genetic linkage analysis; genetic pedigree; genome editing; genome-wide; genomic locus; innovation; insight; malaria infection; malaria transmission; meetings; member; novel; public health relevance; tool; transcriptome sequencing; vector; vector competence; vector mosquito

SUMMARY –The highest global malaria prevalence is in Africa, where the most important vectors are members of the Anopheles gambiae species complex, including the widespread Anopheles coluzzii. Our long-term goal is to dissect the natural genetic differences among mosquitoes that underlie malaria transmission, including differences in malaria susceptibility, ecological adaptation, biting behavior, and insecticide resistance. The objective of this project is to identify the genetic and functional mechanisms of the known major genomic control region for natural A. coluzzii susceptibility to wild P. falciparum. This locus is located in a large region of noncoding DNA on chromosome arm 2L. Frequent alleles in the wild population at this locus strongly influence susceptible or resistant malaria infection outcomes. The central hypothesis is that genetic polymorphism of noncoding regulatory elements explains this locus, and therefore an important fraction of vector genetic variation for P. falciparum infection in nature. The rationale is based on the observation that noncoding genetic polymorphisms control >90% of phenotypic variation in animals, while protein-coding sequence polymorphism contributes little to phenotypic variation. The most important noncoding regulatory elements, enhancers, are responsible for the majority of phenotypic variation. Enhancers are regions ~1 kb in size that modulate target gene expression levels independent of their distance or physical orientation to the targets. Enhancers cannot be predicted by sequence signatures, but require functional assays for detection. In an R21 project, we used a high- throughput functional assay to generate the first comprehensive genome-wide map of enhancers in A. coluzzii, as well as maps of the other noncoding regulatory elements, microRNAs and long noncoding RNAs (miRNAs and lncRNAs). Our specific aims will leverage these new genomic resources to study the influence of natural genetic variation in enhancers and other noncoding regulatory elements for malaria infection outcome: Aim 1) Genetically resolve the major natural locus regulating wild malaria infection; Aim 2) Prioritize the candidate noncoding and coding elements within the locus by testing correlation with infection in a wild mosquito panel; Aim 3) Identify functional mechanisms underlying genetic control of malaria infection. This project is significant because it will determine for the first time the genetic and functional mechanisms underlying the major genomic control region for malaria infection of Anopheles in nature. The proposed project is innovative because the effect of genetic variation of noncoding elements, thought to be the main source of phenotypic variation, has barely been examined in any species, especially in malaria vector mosquitoes, and no mechanism of genetic control over vector competence for Plasmodium in nature is currently known. The results are expected to lead to new malaria control tools rooted in the genome and natural ecology of the vector.

摘要 - 全球最高的疟疾患病率在非洲，最重要的向量是成员 gambiae物种复合物，包括宽度繁殖的山脉coluzzii。我们的长期目标是 剖析疟疾传播基础的蚊子之间的自然遗传差异，包括 疟疾敏感性，生态适应性，咬人行为和杀虫剂耐药性的差异。 该项目的目的是确定已知主要基因组控制的遗传和功能机制 天然A. coluzzii对野生疟原虫的敏感性区域。这个基因座位于一个大区域 染色体ARM 2L上的非编码DNA。该基因座的野生种群中的经常等位基因影响很大 易感或抗性疟疾感染结果。中心假设是 非编码调节元素解释了该基因座，因此是载体遗传变异的重要部分 对于恶性疟原虫感染。基本原理是基于无编码通用的观察结果 多态性控制动物的表型变异的90％，而蛋白质编码序列多态性 对表型变化的贡献很小。最重要的非编码监管元素，增强剂是 负责大多数表型变异。增强剂是调节目标的大小约1 kb的区域 基因表达水平与目标的距离或物理取向无关。增强剂不能 通过序列特征进行预测，但需要功能测定进行检测。在R21项目中，我们使用了高 吞吐量功能评估以生成第一张全面基因组在coluzzii中的综合基因组图， 以及其他非编码调节元件的地图，microRNA和长不编码RNA（miRNA） 和lncrnas）。我们的具体目标将利用这些新的基因组资源来研究自然的影响 疟疾感染结果的增强子和其他非编码调节元素的遗传变异：AIM 1） 基因解决调节野生疟疾感染的主要天然基因座；目标2）优先考虑候选人 通过在野生蚊子面板中测试与感染的相关性，在该基因座内的非编码和编码元素； 目标3）确定疟疾感染遗传控制的基础功能机制。这个项目很重要 因为它将首次确定主要基因组的遗传和功能机制 自然界疟疾感染的疟疾感染区域。拟议的项目具有创新性，因为效果 非编码元件的遗传变异（认为是表型变异的主要来源）几乎没有 在任何物种中，特别是在疟疾载体蚊子中检查了我们，没有遗传控制机制 目前已知自然界的载体能力过多。结果预计将导致新 疟疾控制工具植根于载体的基因组和自然生态学。