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

描述（由申请人提供）：通过在受精卵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）的方法对小鼠进行基因修饰的能力大大增加了小鼠在生物医学研究中的价值和使用。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），对雌性小鼠的术后护理（可能包括使用镇痛药），以及进行手术的人的技术专长。与此同时，动物研究界有一种强烈的愿望，即减少用于研究目的的动物的发病率、压力和痛苦。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。这个过程快速（转移大约需要10秒），不需要手术（包括相关的麻醉和术后护理），并且不需要什么专业知识。我们的初步数据表明，在转基因小鼠的生产中，我们的NSET装置与标准手术胚胎移植在小鼠中同样成功。初步研究采用手工制作的NSET装置进行。我们现在已经制造了一个NSET设备，使我们能够生产大量具有高度均匀性的NSET设备。本提案的目的是证明该制造的NSET装置在非手术移植胚胎方面的功效，(1)可以通过DNA微注射进行转基因，(2)已经冷冻保存，(3)通过冷冻保存的精子体外受精产生的胚胎。这些目标的成功将为NSET设备在学术和生物/制药小鼠设施的商业化提供基础。

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.