A non-surgical embryo transfer (NSET) device for producing gene-modified mice

用于生产基因修饰小鼠的非手术胚胎移植（NSET）装置

• 批准号: 7746604
• 负责人: Shannon Bass
• 金额: $ 10万
• 依托单位: PARATECHS CORP.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-03 至 2010-09-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746604
• 关键词: Analgesics; Anesthesia procedures; Animal Experimentation; Biomedical Research; Caring; Communities; Contracts; DNA; Data; Development; Devices; Embryo; Embryo Transfer; Female; Fertilization in Vitro; Foundations; Gene Targeting; Gene Transfer; Gene-Modified; Genes; Goals; Grant; Hand; Hand functions; Lead; Legal patent; Licensing; Mediating; Medical; Methods; Microinjections; Modification; Molds; Morbidity - disease rate; Mouse Strains; Mus; Operative Surgical Procedures; Pain; Performance; Persons; Phase; Postoperative Period; Procedures; Production; Research; Savings; Scientist; Speed; Stress; Technical Expertise; Technology; Time; Transgenic Mice; Transgenic Organisms; Work; animal pain; base; commercialization; cost; embryonic stem cell; improved; large scale production; novel; public health relevance; sperm cell; success

DESCRIPTION (provided by applicant): The ability to genetically modify mice through methods of gene transfer in fertilized mouse embryos (transgenic mice) or gene modification in embryonic stem (ES) cells (gene targeting) has dramatically increased the value and use of mice in biomedical research. . The production of gene-modified mice requires the surgical transfer of embryos into female mice. This surgery requires anesthesia (with potential complications), post-operative care of female mice (which may include the use of analgesics), as well as technical expertise on the part of the person carrying out the surgery, as. At the same time, there is a strong desire in the animal research community to reduce the morbidity, stress and pain of animals used for research purposes. To overcome these problems, we have developed a Non-Surgical Embryo Transfer (NSET) device that can be used for the non-surgical transfer of embryos into female mice. This procedure is rapid (transfer requires about 10 seconds), does not require surgery (including the associated anesthesia and post-operative care), and requires little expertise. Our preliminary data indicate that our NSET device is equally successful as standard surgical embryo transfer in mice in the production of transgenic mice. Preliminary studies were performed with hand-made NSET devices. We have now manufactured a NSET device, enabling us to produce large numbers of NSET devices with a high degree of uniformity. The aims of this proposal are to demonstrate the efficacy of this manufactured NSET device for the non-surgical transfer of embryos that (1) have may be transgenic by DNA microinjection, (2) have been cryopreserved, and (3) were generated by in vitro fertilization of embryos with cryopreserved sperm. Success in these aims will provide the foundation for the commercialization of the NSET device for academic and bio/pharma mouse facilities. PUBLIC HEALTH RELEVANCE: We have developed a device for the non-surgical transfer of embryos into recipient female mice that represents a substantial advance over existing methods of generating gene-modified mice in that it eliminates surgery, including anesthesia and post-operative care. The goal of this proposal is to determine the efficacy of a manufactured non-surgical embryo transfer device for the transfer of embryos that are transgenic, cryopreserved, or generated by in vitro fertilization.

描述（由申请人提供）：通过在受精小鼠胚胎中进行基因转移（转基因小鼠）或在胚胎干（ES）细胞中进行基因修饰（基因靶向）的方法对小鼠进行遗传修饰的能力显著增加了小鼠在生物医学研究中的价值和用途。.基因改造小鼠的生产需要将胚胎移植到雌性小鼠体内。这种手术需要麻醉（有潜在的并发症），雌性小鼠的术后护理（可能包括使用止痛药），以及进行手术的人的技术专长。同时，动物研究界强烈希望减少用于研究目的的动物的发病率、压力和疼痛。为了克服这些问题，我们开发了一种非手术胚胎移植（NSET）装置，可用于将胚胎非手术移植到雌性小鼠体内。这个过程是快速的（转移需要大约10秒），不需要手术（包括相关的麻醉和术后护理），并且需要很少的专业知识。我们的初步数据表明，我们的NSET装置在转基因小鼠的生产中与小鼠中的标准外科胚胎移植同样成功。用手工制作的NSET器械进行了初步研究。我们现在已经制造了NSET器件，使我们能够生产大量具有高度一致性的NSET器件。本提案的目的是证明该生产的NSET器械用于非手术移植胚胎的有效性，这些胚胎（1）可能通过DNA显微注射转基因，（2）已经冷冻保存，（3）通过体外受精胚胎与冷冻保存的精子产生。这些目标的成功将为学术和生物/制药小鼠设施的NSET设备的商业化提供基础。 公共卫生关系：我们已经开发出一种将胚胎非手术移植到受体雌性小鼠中的装置，该装置代表了对现有产生基因修饰小鼠的方法的实质性进步，因为它消除了手术，包括麻醉和术后护理。本提案的目的是确定制造的非手术胚胎移植装置用于移植转基因、冷冻保存或体外受精产生的胚胎的有效性。

项目成果

A rapid and effective nonsurgical artificial insemination protocol using the NSET™ device for sperm transfer in mice without anesthesia.

• DOI: 10.1007/s11248-015-9887-3
• 发表时间: 2015-08
• 期刊: Transgenic research
• 影响因子: 3
• 作者: Stone BJ;Steele KH;Fath-Goodin A
• 通讯作者: Fath-Goodin A

Nonsurgical Embryo Transfer Protocol for Use with the NSET™ Device.

适用于 NSET™ 设备的非手术胚胎移植协议。

• DOI: 10.1007/978-1-4939-9837-1_9
• 发表时间: 2020
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Stone,BarbaraJ