Tools for regulated gene expression in Entamoeba invadens

内阿米巴入侵基因表达调控工具

基本信息

批准号：
8998927
负责人：
UPINDER SINGH
金额：
$ 20.06万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8998927
关键词：
Amoeba genus Biological Models Biological Process Biology Cause of Death Cells Communities Cyst Data Development Developmental Biology Disease Dissection Dominant-Negative Mutation Dysentery Entamoeba Entamoeba histolytica Entamoeba invadens Etiology Exploratory/Developmental Grant Funding Gene Expression Gene Proteins Gene Silencing Generations Genetic Genetic study Genome Genomics Goals Grant Health Human In Vitro Knowledge Life Cycle Stages Liver Abscess Messenger RNA Methods Molecular Organism Parasites Pathogenesis Pathway interactions Phenotype Ploidies Process Proteins Protocols documentation Publications Publishing RNA Interference Reagent Regulation Role Small RNA Staging Syndrome System Technology Transfer Tetanus Helper Peptide Tissues Transfection Work base disease transmission enteric pathogen excystation experience gene replacement genetic manipulation interest metabolomics method development pathogen prevent promoter protein expression routine practice success tool transcriptome transmission process vector

项目摘要

DESCRIPTION (provided by applicant): Entamoeba histolytica, a protozoan parasite, is an important human pathogen. Diseases caused by E. histolytica include dysentery and liver abscesses, and this organism is a leading parasitic cause of death on a global scale. The life cycle of Entamoeba involves stage inter-conversion between trophozoites and cysts. Importantly, the conversion to cysts allows the pathogen to disseminate to new hosts while development to trophozoites allows the organism to cause invasive disease in the host. Thus, stage interconversion is essential for disease transmission and pathogenesis. However, despite being central to amebic biology, stage conversion is poorly understood. Many factors have contributed to the paucity of data including the inability to reproduce the developmental cycle in E. histolytica. Instead, the reptilian ameba E. invadens, which has the same disease pathogenesis as E. histolytica, has been used as a model system to study Entamoeba development. In E. invadens, both encystation and excystation can be recapitulated with high efficiency in vitro. However, until recently a poorly annotated genome and lack of methods for genetic manipulation have prevented full exploitation of this system. The recent re-sequencing of the E. invadens genome, publication of the transcriptome of stage conversion, and development of methods for constitutive gene expression open up new avenues for discovery. We aim to build upon these recent successes by establishing new tools for the study of E. invadens including (i) regulated gene expression, (ii) gene silencing, and (iii) development of negative selection and single cell markers. These methods will make possible new experimental strategies for studying Entamoeba stage conversion, and greatly aid the entire Entamoeba community. Dissecting the developmental cascade is crucial to understanding the biology of this important human pathogen.

描述（由适用提供）：原生动物寄生虫Entamoeba Histolictica是一种重要的人类病原体。由E. astolytica引起的疾病包括痢疾和肝脓肿，这种生物是全球范围内的主要寄生虫死亡原因。 Entamoeba的生命周期涉及滋养体和囊肿之间的阶段相互转化。重要的是，向囊肿的转化使病原体可以传播到新宿主，而发展为滋养体可以使生物体在宿主中引起侵入性疾病。这是阶段互连对于疾病传播和发病机理至关重要的。然而，尽管对阿米比克生物学是核心，但阶段转换知之甚少。许多因素导致数据缺乏，包括无法再现组织溶液性的发育周期。取而代之的是，具有与Histolytica E. histolictica相同的疾病发病机理的复制品Ameba E. Invadens已被用作研究肠发展的模型系统。在E. Inscadens中，可以通过体外高效率概括百科全书和外观。然而，直到最近，注释不足的基因组和缺乏基因操纵方法还阻止了该系统的完全开发。最近对E. Is Inted Genome，阶段转换的转录组的发表以及构成基因表达方法的发展开发了新的发现途径的方法。我们的目的是通过建立用于研究E.侵袭性的新工具，包括（i）（i）调节基因表达，（ii）基因沉默和（iii）阴性选择和单细胞标记物的发展。这些方法将为研究Entamoeba舞台转换而成为可能的新实验策略，并极大地帮助整个Entamoeba社区。解剖发育级联对于理解这种重要人类病原体的生物学至关重要。