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The use of lentiviral vectors to obtain transgenic rats.

Methods in molecular biology (Clifton, N.J.) (2009-12-17)
Séverine Remy, Tuan Huy Nguyen, Séverine Ménoret, Laurent Tesson, Claire Usal, Ignacio Anegon
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hyaluronidase from bovine testes, Type IV-S, powder, suitable for mouse embryo cell culture, 750-3000 units/mg solid
Pricing and availability is not currently available.
Sigma-Aldrich
Amplifluor® UniPrimer II Fluorescein
Pricing and availability is not currently available.