PROPAGATION OF MONKEY MODELS OF HUMAN DISEASE

人类疾病的猴子模型的传播

• 批准号: 6639899
• 负责人: DON P WOLF
• 金额: $ 41.2万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6639899
• 关键词: AIDS; AIDS vaccines; HIV infections; MHC class I antigen; Macaca mulatta; alleles; animal breeding; cryopreservation; disease /disorder model; egg /ovum; embryo /fetus culture; fertility; genotype; heterozygote; homozygote; human immunodeficiency virus; model design /development; monozygotic twins; simian immunodeficiency virus; sperm; vaccine development

DESCRIPTION (Adapted from the applicant's abstract): There is currently a significant need for populations of animals with specified genotypes which cannot be satisfied by the importation of animals from the wild or by the identification and propagation of valuable founder animals by selective breeding. Indian-origin, rhesus macaques carrying the MHC class 1 allele, A*01, are particularly needed for vaccine development research. The objective of this application is to demonstrate that ARTs can be applied to the rapid, efficient propagation of a valuable founder animal and to the production of identical twins using existing technology, thereby establishing a new approach to satisfying animal requirements of the biomedical research community. The rationale for focusing on a homozygous, founder male is simple. All offspring produced by this animal will be Mamu-A*01 positive. Moreover, if heterozygous carriers of the A*01 allele are used as oocyte donors, 50% of the offspring will be homozygous for the allele, creating additional founder animals. The applicants have access to a male at the NERPRC that is presumed homozygous for the Mamu-A*01 allele based on the fact that 100% (15 of 15; K. Mansfield, personal communication) of his offspring are Mamu-A*01 positive. The number of sperm in an average ejaculate, when used in conjunction with ICSI, can result in the production of hundreds, if not thousands, of embryos. Sperm collection, preservation during transportation or storage and use of ICSI is established technology in the applicants' laboratory, allowing a high probability of success. The investigators propose to establish a new paradigm for the efficient, cost effective propagation of founder monkeys and populations of valuable, genetically defined macaques, including identical twins, through conduction of the following specific aims: 1) Produce 25 Mamu-A*01 positive animals in the first year and 50 animals per year for the duration of this study using sperm from a homozygous Mamu-A*01 positive donor. This aim will establish the paradigm and create new populations of heterozygous and homozygous animals without impacting the natural reproduction of either the sperm or the oocyte donors. The applicants will confirm homozygosity in candidate animals by creating embryos with their gametes and monitoring the presence or absence of the Mamu-A*01 allele in these embryos; and 2) Employ blastomere separation and culture or embryo splitting technology to increase embryo numbers and to produce genetically identical animals. The applicants will select the optimal approach to twinning in year 1 and produce 5 sets of identical twins per year during years 2-5 of the study.

描述（改编自申请人的摘要）：目前有一个 显著需要具有特定基因型的动物群体， 不能通过从野生动物进口或 通过选择性的方法鉴定和繁殖有价值的创始动物 育种 印度裔恒河猴携带MHC 1类等位基因， A*01，特别需要用于疫苗开发研究。 客观 本申请的目的是证明ART可以应用于快速， 有效繁殖有价值的创始人动物和生产 同卵双胞胎使用现有技术，从而建立一种新的方法， 来满足生物医学研究界对动物的要求。 的 关注纯合的创始男性的基本原理很简单。 所有幼鼠 Mamu-A*01阳性。 此外，如果杂合子 A*01等位基因携带者被用作卵母细胞供体，50%的后代 将是等位基因的纯合子，产生额外的创始动物。 的 申请人可以在NERPRC获得一名男性，该男性被认为是纯合子， Mamu-A*01等位基因是基于100%（15/15; K.曼斯菲尔德， 他的后代的个人通信）是Mamu-A*01阳性。 数量 当与ICSI结合使用时， 导致产生数百甚至数千个胚胎。 精子 ICSI的收集、运输或储存过程中的保存和使用是 在申请人的实验室建立的技术，允许高 成功的概率。 研究人员建议建立一个新的范式， 有效地繁殖创始猴和有价值的， 基因定义的猕猴，包括同卵双胞胎，通过传导， 具体目标如下：1）在实验室中生产25只Mamu-A*01阳性动物 第一年和本研究期间每年50只动物使用精子 来自纯合子Mamu-A*01阳性供体。 这一目标将确立 并创造新的杂合和纯合动物种群 而不影响精子或卵母细胞的自然繁殖 捐助者。 申请人将通过以下方式确认候选动物的纯合性： 用它们的配子创造胚胎， 这些胚胎中的Mamu-A*01等位基因;和2）采用卵裂球分离， 培养或胚胎分裂技术，以增加胚胎数量， 产生基因相同的动物。 申请人将选择最佳的 在第一年接近双胞胎，每年产生5对同卵双胞胎 在研究的第2-5年。