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开发疫苗和研究大多数人类疾病至关重要。现代啮齿类动物研究越来越强大，有能力操纵小鼠和大鼠的基因组，使它们接近模拟复杂的人类疾病，如阿尔茨海默氏症和动脉硬化。培育转基因小鼠和大鼠需要将研究人员操纵的胚胎转移到雌性小鼠受体中，在那里它们可以完成发育。到目前为止，这些移植都需要将胚胎植入子宫的外科手术。最近，已经开发出一种装置，使这些胚胎移植无需手术即可进行，从而消除了术后恢复期，从而减少了动物的痛苦。本提案通过建立胚胎移植的最佳年龄和小鼠品系（Aim 1），确定胚胎干细胞是否可以有效和高效地使用该装置移植（Aim 2），确定该装置是否适用于人工授精（Aim 3），研究并扩展了该胚胎移植装置的实用性。直接观察子宫内的装置，这可能有助于设计改进（目标4），并量化与手术过程相关的压力测量，以支持该过程在研究界的广泛采用（目标5）。该项目的主要目标是产生数据来证明这种胚胎移植装置的有效性，从而使研究人员能够用一种同样有效的非手术方法取代外科手术。证明这种非手术移植方法在一系列条件下是有效的，将扩大其应用范围，从而最大限度地提高其影响，使研究人员能够改进和减少在小鼠身上进行的外科手术的数量。

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.