RIKEN BioResource Research Center, Tsukuba, Ibaraki, Japan.
Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba, Ibaraki, Japan.
Methods Mol Biol. 2023;2637:247-254. doi: 10.1007/978-1-0716-3016-7_19.
The golden (Syrian) hamster (Mesocricetus auratus) is a small rodent belonging to the Cricetidae family. Golden hamsters have several unique characteristics that are advantageous in the study of reproductive and developmental biology: a highly stable 4-day estrous cycle, a high responsiveness to conventional superovulation methods, and a shortest gestation period (16 days) known among eutherian mammals. Besides these advantages, the technical ease of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in this species has contributed much to our understanding of the basic mechanisms of mammalian fertilization. However, the exceptionally strong in vitro developmental block of hamster embryos, especially at the two-cell stage, has hampered the production of genetically modified hamsters, which has resulted in limited use of this species for biomedical research. However, the recently developed in vivo genome editing method (improved genome editing via oviductal nucleic acid delivery, i-GONAD) has overcome this shortcoming and made production of gene-edited hamsters much easier than before. This method has the potential to provide a means of reexamining genes whose functions cannot be identified using mouse models, thus leading to the better understanding of gene functions in mammals. In this chapter, we present our procedure for editing the genome of the golden hamster using i-GONAD.
金黄仓鼠(Mesocricetus auratus)是一种小型啮齿动物,属于仓鼠科。金黄仓鼠具有几个独特的特点,在生殖和发育生物学的研究中具有优势:高度稳定的 4 天发情周期,对常规超数排卵方法的高度反应性,以及在真兽类哺乳动物中最短的妊娠期(16 天)。除了这些优势外,该物种中体外受精(IVF)和胞质内精子注射(ICSI)的技术简便性极大地促进了我们对哺乳动物受精基本机制的理解。然而,仓鼠胚胎在体外发育的异常强烈阻断,特别是在二细胞阶段,阻碍了基因修饰仓鼠的生产,这限制了该物种在生物医学研究中的应用。然而,最近开发的体内基因组编辑方法(通过输卵管核酸传递的改良基因组编辑,i-GONAD)克服了这一缺点,使基因编辑仓鼠的生产比以前更容易。这种方法有可能提供一种重新研究使用小鼠模型无法确定其功能的基因的手段,从而更好地理解哺乳动物中的基因功能。在本章中,我们介绍了使用 i-GONAD 编辑金黄仓鼠基因组的程序。