Genomic Engineering in Drosophila

果蝇基因组工程

• 批准号: 7746473
• 负责人: Yang Hong
• 金额: $ 17.04万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-12 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746473
• 关键词: Address; Alleles; Animal Model; Back; Bacteriophages; Biochemical; Biological; Biological Assay; Biological Models; Cell Polarity; Cells; Chimeric Proteins; Chromosomes, Human, Pair 3; Complex; DNA; DNA Integration; DNA Structure; Development; Disease Pathway; Disease model; Drosophila genome; Drosophila genus; Effectiveness; Engineering; Essential Genes; Frequencies; Gene Targeting; Genes; Genetic; Genetic Models; Genetic Recombination; Genome; Genomics; Goals; Homologous Gene; Human Genetics; In Vitro; Integrase; Knock-out; Left; Lesion; Location; Mediating; Modification; Point Mutation; Positioning Attribute; Process; Research Personnel; Scheme; Site; Site-Directed Mutagenesis; Structure; System; Testing; Transgenic Organisms; Work; arm; base; design; genetic manipulation; homologous recombination; human disease; in vivo; mutant; novel; public health relevance; recombinase; research study; tool; vector

DESCRIPTION (provided by applicant): In this proposal we describe the development of a novel genetic manipulation system termed "genomic engineering" that will permit efficient, unlimited and directed modification of a chosen genomic locus in Drosophila. Genomic engineering is a two-step process. First, through an optimized gene targeting routine, a founder knock-out line is generated by deleting the target gene and replacing it with a small DNA recombination site of phage integrase FC31. Second, DNA integration by FC31 is used to reintroduce modified target gene DNA into the native locus in the founder knock-out line. Since FC31 mediated DNA integration is highly efficient and non-discriminating, virtually any desired genetic modification can be generated with high throughput efficiency. Genomic engineering will have a profound and revolutionary impact as a tool for directed engineering of the Drosophila genome. First, genomic engineering overcomes the inherent inefficiency and limitations of homologous recombination. It provides a virtually unlimited approach to generate any desired mutant allele of a target gene for genetic, biochemical and cell biologic assays. Second, for constructing useful and informative human disease models, genomic engineering makes it possible to precisely replicate the human genetic lesions into the Drosophila homologous genes. Finally, by systematically generating founder knock-out lines for conserved and essential genes in the long term, we will transform Drosophila into a far more efficient and versatile genetic model organism that will also be vastly more accessible and attractive to non-fly researchers for addressing their biological questions. PUBLIC HEALTH RELEVANCE: Drosophila is a leading genetic model system for addressing crucial biological questions in human diseases. Genomic engineering is a new powerful genetic tool that will significantly facilitate the analysis of complex disease pathways and the construction of better disease models in Drosophila.

描述（由申请人提供）：在本提案中，我们描述了一种称为"基因组工程"的新型遗传操作系统的开发，该系统将允许对果蝇中选定的基因组位点进行有效、无限制和定向的修饰。基因组工程是一个两步的过程。首先，通过优化的基因靶向程序，通过删除靶基因并用噬菌体整合酶FC 31的小DNA重组位点替换它来产生创始人敲除系。第二，通过FC 31的DNA整合用于将修饰的靶基因DNA重新引入创始者敲除系中的天然基因座中。由于FC 31介导的DNA整合是高效和非鉴别性的，因此几乎可以以高通量效率产生任何所需的遗传修饰。基因组工程作为果蝇基因组定向工程的工具，将产生深远而革命性的影响。首先，基因组工程克服了同源重组固有的低效率和局限性。它提供了一种几乎不受限制的方法来产生用于遗传、生物化学和细胞生物学测定的靶基因的任何所需突变等位基因。第二，为了构建有用的和信息丰富的人类疾病模型，基因组工程使得精确地将人类遗传病变复制到果蝇同源基因中成为可能。最后，通过长期系统地产生保守和必需基因的创始人敲除系，我们将把果蝇转化为一种更有效和更通用的遗传模式生物，这也将大大提高非飞行研究人员的可访问性和吸引力，以解决他们的生物学问题。公共卫生相关性：果蝇是解决人类疾病中关键生物学问题的领先遗传模型系统。基因组工程是一种新的强大的遗传工具，将显着促进复杂的疾病途径的分析和更好的疾病模型在果蝇中的构建。

项目成果

W::Neo: a novel dual-selection marker for high efficiency gene targeting in Drosophila.

W::Neo：一种新型双选择标记，用于果蝇高效基因靶向。

• DOI: 10.1371/journal.pone.0031997
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Zhou,Wenke;Huang,Juan;Watson,AnnieM;Hong,Yang
• 通讯作者: Hong,Yang

A conserved polybasic domain mediates plasma membrane targeting of Lgl and its regulation by hypoxia.

• DOI: 10.1083/jcb.201503067
• 发表时间: 2015-10-26
• 期刊: The Journal of cell biology
• 作者: Dong W;Zhang X;Liu W;Chen YJ;Huang J;Austin E;Celotto AM;Jiang WZ;Palladino MJ;Jiang Y;Hammond GR;Hong Y
• 通讯作者: Hong Y