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

描述(由申请人提供)：在研究中使用研究动物对于疫苗的开发和对大多数人类疾病的研究都是必不可少的。现代啮齿动物研究越来越强大，有能力操纵小鼠和大鼠的基因组，使它们接近模拟阿尔茨海默氏症和动脉粥样硬化等复杂的人类疾病。要产生转基因小鼠和大鼠，需要将研究人员操纵的胚胎转移到受体雌性小鼠身上，在那里它们可以完成发育。到目前为止，这些移植需要将胚胎植入子宫的外科手术。最近，已经开发出一种设备，可以在不进行手术的情况下进行胚胎移植，从而消除了手术后恢复期，从而减轻了动物的痛苦。这项建议通过确定胚胎移植的最佳年龄和小鼠品系(目标1)，确定胚胎干细胞能否有效和高效地通过该设备转移(目标2)，确定该设备是否可用于人工授精(目标3)，在子宫内直接观察该设备以改进设计(目标4)，并量化与外科手术相关的压力测量，以支持在研究界广泛采用这一程序(目标5)，从而调查和扩大这种胚胎移植设备的用途。该项目的主要目标是提供数据，证明这种胚胎移植设备的有效性，从而使研究人员能够用同样有效的非手术方法取代外科手术。证明这种非手术转移方法在一系列条件下是有效的，将扩大它在这一范围内的应用，从而最大限度地发挥其影响，使研究人员能够改进和减少在小鼠身上进行的手术程序的数量。 公共卫生相关性：动物在研究中的使用对于生物医学研究和开发用于研究的基因工程小鼠的能力是必不可少的；这些动物正变得更加重要，因为它们可以被改造成模仿大多数人类疾病。小鼠的基因工程需要，作为许多其他事情之一，将胚胎移植到小鼠体内，在那里它们完成发育。该项目支持进一步开发一种设备，使这些胚胎移植能够在不进行手术的情况下进行，从而大大减少这些研究动物经历的疼痛类别，从而支持生物医学的进步。

DESCRIPTION (provided by applicant): The use of research animals in research has been essential to development of vaccines and study of most human diseases. Modern rodent research is increasingly powerful with the ability to manipulate the genomes of mice and rats such that they closely mimic complex human diseases such as Alzheimer's and artherosclerosis. Generating transgenic mice and rats, requires that embryos that are manipulated by researchers be transferred into recipient female mice where they can complete their development. Up to this point, these transfers have required surgical procedures in which the embryo is implanted into the uterus. Recently, a device has been developed that enables these embryo transfers to be performed without surgery which eliminates the post-operative recovery period thereby reducing pain of the animals. This proposal investigates and expands upon the utility of this embryo transfer device by establishing the optimal ages and mouse strains for embryo transfer (Aim 1), determining whether embryonic stem cells can be effectively and efficiently transferred with this device (Aim 2), determining whether the device is useful for artificial inseminations (Aim 3), directly visualizing the device in the uterus which may enable design improvements (Aim 4) and quantifying measures of stress relative to surgical procedures to support widespread adoption of this procedure in the research community (Aim 5). The primary project objective is to produce data that will prove the efficacy of this embryo transfer device and thereby enable researchers to replace surgical procedures with a non-surgical method that is equally effective. Proving that this non-surgical transfer method is effective under a range of conditions will expand its use over this range of applications and thereby maximize its impact in enabling researchers to refine and reduce the number of surgical procedures performed in mice. PUBLIC HEALTH RELEVANCE: Animal use in research is essential for biomedical research and with the developing abilities to genetically engineer mice for research; these animals are becoming even more important as they can be modified to mimic most human diseases. Genetic engineering of mice requires, as one of many other things, the ability to transfer embryos into mice where they complete their development. This project supports further development of a device that enables these embryo transfers to be performed without surgery and therefore greatly reduces the pain category that these research animals experience and thereby supports progress in biomedicine.