Selfish gene and genetic control of vector-borne diseases

自私基因与媒介传播疾病的遗传控制

• 批准号: 7364642
• 负责人: Zach N. Adelman
• 金额: $ 21.79万
• 依托单位: VIRGINIA POLYTECHNIC INST AND ST UNIV
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7364642
• 关键词: Aedes; African; Asians; Bacteria; Bacterial Transformation; Biological Assay; Cells; Cessation of life; Chagas Disease; Cleaved cell; Competence; Complement; Culicidae; Cultured Cells; DNA; DNA Transposable Elements; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; Development; Disadvantaged; Disease Vectors; Drosophila genus; Elements; Embryo; Escherichia coli; Event; Excision; Eye; Foundations; Funding; Future; Generations; Genes; Genetic; Genome; Germ Cells; Homing; Human; In Vitro; Individual; Injection of therapeutic agent; Investigation; Investments; Life; Ligation; Malaria; Methods; Modification; Molecular Analysis; Molecular Weight; Nature; Outcome; Parasites; Pigmentation physiologic function; Pigments; Plasmids; Population; Proteins; Public Health; Rate; Reporting; Selfish Genes; Site; Source; Sucrose; System; Target Populations; Technology; Transfection; Transgenes; Trypanosoma cruzi; Vector-transmitted infectious disease; Work; Yellow Fever; base; cinnabar; design; endonuclease; human disease; improved; neglect; pathogen; plasmid DNA; research study; transmission process; vector; vector mosquito

DESCRIPTION (provided by applicant): This proposal aims to determine the potential for selfish genes, AKA homing endonucleases, to serve as a method of dispersing and fixing genes into natural mosquito populations that reduce the vector competence of mosquitoes to transmit pathogens, such as the causative agents of dengue fever and malaria. In the first aim of this proposal we seek to determine the ability of various homing endonucleases to generate dsDNA breaks using a two-plasmid based assay. This assay has been designed so that successful cutting by the homing endonuclease will result in the loss of a negative selectable marker. Plasmid-based assays will be conducted in both cultured mosquito cells as well as in live embryos. In the second aim we seek to determine the rate at which homing endonucleases can excise a transgene from the mosquito genome. A marker gene flanked by homing endonuclease recognition sites will be inserted into the mosquito genome, followed by the introduction of a homing endonuclease which should excise the marker. Homing endonucleases found to be functional in both plasmid-based assays and transgene excision assays will serve as the foundation for future work at adapting homing endonucleases as a method of gene drive. Public health relevance: This work is part of a larger, ongoing strategy to control human pathogens using genetics. Such a genetic control strategy is founded on the hypothesis that reduced/ablated vector competence will result in a corresponding reduction/elimination of human disease, and is based on three elements: the development of anti-pathogen effector genes, the ability to introduce effector genes into mosquito vectors, and the ability to spread these effector genes into wild populations. The first two elements have seen substantial progress in the past decade, and this proposal is designed complement this work so genetic control strategies can become a reality in effecting and improving public health outcomes due to vector-borne diseases.

描述（由申请人提供）：本提案旨在确定自私基因（AKA归巢核酸内切酶）作为将基因分散和固定到天然蚊子种群中的方法的潜力，该方法降低蚊子传播病原体（如登革热和疟疾的病原体）的载体能力。在该提议的第一个目的中，我们试图使用基于双质粒的测定来确定各种归巢核酸内切酶产生dsDNA断裂的能力。该测定被设计成使得归巢核酸内切酶的成功切割将导致阴性选择标记的损失。将在培养的蚊子细胞以及活胚胎中进行基于质粒的测定。在第二个目标中，我们试图确定归巢核酸内切酶可以从蚊子基因组中切除转基因的速率。将侧翼为归巢核酸内切酶识别位点的标记基因插入蚊子基因组中，然后引入归巢核酸内切酶，该核酸内切酶应切除标记。在基于质粒的测定和转基因切除测定中发现有功能的归巢核酸内切酶将作为未来工作的基础，以使归巢核酸内切酶适应作为基因驱动的方法。公共卫生相关性：这项工作是利用遗传学控制人类病原体的更大的持续战略的一部分。这种遗传控制策略是建立在这样的假设上的，即降低/消除的载体能力将导致相应的减少/消除人类疾病，并且基于三个要素：抗病原体效应基因的发展，将效应基因引入蚊子载体的能力，以及将这些效应基因传播到野生种群中的能力。前两个要素在过去十年中取得了重大进展，本提案旨在补充这项工作，以便遗传控制战略能够在影响和改善病媒传播疾病造成的公共卫生结果方面成为现实。

项目成果

Homing endonucleases catalyze double-stranded DNA breaks and somatic transgene excision in Aedes aegypti.

• DOI: 10.1111/j.1365-2583.2009.00905.x
• 发表时间: 2009-10
• 期刊: Insect molecular biology
• 影响因子: 2.6
• 作者: Traver BE;Anderson MA;Adelman ZN
• 通讯作者: Adelman ZN

Robust heat-inducible gene expression by two endogenous hsp70-derived promoters in transgenic Aedes aegypti.

• DOI: 10.1111/j.1365-2583.2011.01116.x
• 发表时间: 2012-02
• 期刊: Insect molecular biology
• 影响因子: 2.6
• 作者: Carpenetti TL;Aryan A;Myles KM;Adelman ZN
• 通讯作者: Adelman ZN

Validation of novel promoter sequences derived from two endogenous ubiquitin genes in transgenic Aedes aegypti.

• DOI: 10.1111/j.1365-2583.2010.01005.x
• 发表时间: 2010-08
• 期刊: Insect molecular biology
• 影响因子: 2.6
• 作者: Anderson MA;Gross TL;Myles KM;Adelman ZN
• 通讯作者: Adelman ZN

Identification and characterization of heat shock 70 genes in Aedes aegypti (Diptera: Culicidae).

• DOI: 10.1603/033.046.0313
• 发表时间: 2009-05
• 期刊: Journal of medical entomology
• 影响因子: 2.1
• 作者: Gross TL;Myles KM;Adelman ZN
• 通讯作者: Adelman ZN

Germline excision of transgenes in Aedes aegypti by homing endonucleases.

• DOI: 10.1038/srep01603
• 发表时间: 2013
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Aryan, Azadeh;Anderson, Michelle A. E.;Myles, Kevin M.;Adelman, Zach N.
• 通讯作者: Adelman, Zach N.