Remy Séverine, Nguyen Tuan Huy, Ménoret Séverine, Tesson Laurent, Usal Claire, Anegon Ignacio
INSERM, UMR643, Nantes, France.
Methods Mol Biol. 2010;597:109-25. doi: 10.1007/978-1-60327-389-3_8.
Lentiviral vectors are now well recognized as good vehicles for gene delivery. This is because they can efficiently transduce both dividing and post-mitotic cells, and stably integrate into the host genome allowing for long-term expression of the transgene. Their potential utility for the generation of transgenic animals has been recognized as an attractive and promising alternative to the conventional DNA-microinjection method which lacks efficiency. The initial success of lentiviral transgenesis in mice considerably broadened its use in other species, in which classical transgenic techniques are difficult, such as in the rat.In this chapter, we describe detailed procedures for both the production of human immunodeficiency virus-1 (HIV-1)-derived lentiviral vectors and for the generation of transgenic rats by injection of these vectors into the perivitelline space of fertilized one-cell eggs.
慢病毒载体如今已被公认为是基因传递的优良载体。这是因为它们能够有效地转导分裂细胞和有丝分裂后的细胞，并稳定整合到宿主基因组中，从而使转基因能够长期表达。它们在生成转基因动物方面的潜在用途，已被视为一种有吸引力且前景广阔的替代方法，可替代缺乏效率的传统DNA显微注射法。慢病毒转基因技术在小鼠中的初步成功极大地拓展了其在其他物种中的应用，比如在大鼠中，传统转基因技术很难实施。在本章中，我们将详细描述源自人类免疫缺陷病毒1型（HIV-1）的慢病毒载体的生产过程，以及通过将这些载体注射到单细胞受精卵的卵周隙中来生成转基因大鼠的过程。
