Microinjection Station for Utah Transgenic and Gene Targeting Mouse Core

描述（由申请人提供）：我们建议购买蔡司显微注射站用于犹他大学转基因和基因靶向小鼠核心。这个核心设施被NIH资助的研究人员大量使用，并为他们发表的许多手稿做出了贡献。该仪器及其相关硬件的功能包括将小鼠胚胎干细胞注射到囊胚中，将DNA注射到受精卵的原核中，8细胞期胚胎干细胞注射（ES）细胞，辅助体外受精（AIVF）和胞浆内单精子注射（ICSI）。许多用于产生转基因小鼠的程序都是在犹他大学开发的，我们拥有最古老的核心设施之一。缺点是我们的显微注射仪器现在已经有20年的历史了，缺乏许多高效小鼠生成所必需的功能。通过原核注射生产基因靶向小鼠和转基因小鼠是哺乳动物研究的重要组成部分。由于需要技术专长，基因靶向和转基因小鼠的产生通常在像我们这样的核心设施中进行。拥有最新的、最先进的设备，每天都在核心中使用，极大地提高了研究人员项目的成功和效率。大多数基因靶向小鼠是通过将靶向胚胎干细胞微注射到囊胚期胚胎中产生的。这是通过强大的显微镜和微操作器来选择并将单个胚胎干细胞注射到每个囊胚中。通过手术将注射的囊胚植入假孕的雌性体内，由雌性携带胚胎至足月。出生的幼崽是嵌合体，可以将目标基因传递给后代。100%种系基因靶向小鼠也可以通过将胚胎干细胞注射到8细胞阶段的胚胎中产生。优秀的光学和微操纵器是必不可少的这些程序。转基因小鼠是通过向受精卵的原核中注射经过处理的DNA而产生的。DNA随机整合到基因组中，如果使用组织特异性启动子来驱动或过度表达感兴趣的基因或报告基因，则很有用。AIVF（辅助体外受精）是通过使用激光在未受精卵的透明带钻孔来实现的，这使得精子更容易与卵子受精，极大地改善了结果。卵胞浆内单精子注射（ICSI）是将固定的精子直接注射到卵子中。如果正常的IVF或AIVF不成功，则使用此程序。在此过程中，进样器必须具有良好的光学性能。

DESCRIPTION (provided by applicant): We propose to purchase a Zeiss microinjection station for use in the University of Utah Transgenic and Gene Targeting Mouse Core. This core facility is heavily used by NIH- funded investigators, and has contributed to many of their published manuscripts. The capabilities of the instrument and its associated hardware include injection of mouse embryonic stem cells into blastocysts, injection of DNA into the pronuclei of fertilized eggs, 8-cell stage injections of embryonic stem (ES) cells, assisted in vitro fertilization (AIVF), and intracytoplasmic sperm injection (ICSI). Many of the procedures used to generate transgenic mice were developed at the University of Utah, and we have one of the oldest core facilities. The downside is that our microinjection instrument is now 20 years old, and lacks many features essential for highly efficient mouse generation. Production of gene-targeted mice and the generation of transgenic mice via pronuclear injection are essential parts of mammalian research. Because of the technical expertise required, the generation of gene targeted and transgenic mice is usually performed in a Core facility such as ours. Having updated, state-of-the-art equipment that is used every day in the Core immensely increases the success and efficiency of projects for researchers. Most gene-targeted mice are generated by microinjecting targeted embryonic stem cells into blastocyst stage embryos. This is done using a powerful microscope and micromanipulators to select and inject individual ES cells into each blastocyst. Injected blastocysts are surgically implanted into pseudopregnant females which carry the embryos to term. Pups born are chimeras which can pass on the targeted gene to its offspring. 100% germline gene-targeted mice can also result from injecting ES cells into 8-cell stage embryos. Excellent optics and micromanipulators are essential to these procedures. Transgenic mice are produced by injecting manipulated DNA into the pronucleus of fertilized eggs. DNA integrates into the genome randomly and is useful if using a tissue-specific promoter to drive or overexpress a gene of interest or reporter. AIVF (assisted in vitro fertilization) is performed by boring a hole in the zona pellucida of an unfertilized egg using a laser, which makes it easier for sperm to fertilize the egg, immensely improving results. ICSI (intracytoplasmic sperm injection) is used to directly inject immobile sperm into an egg. This procedure is used if normal IVF or AIVF is unsuccessful. It is imperative that the injector have excellent optics in this procedure.