Generation of transgenic cattle by lentiviral gene transfer into oocytes.
The potential benefits of transgenic cattle range from the production of large quantities of pharmaceutically relevant proteins to agricultural improvement. However, the production of transgenic cattle is presently time-consuming and expensive because of the inefficiency of the classical DNA microinjection technique. Here, we report the use of lentiviruses for the efficient generation of transgenic cattle. Initial attempts to produce transgenic cattle by lentiviral infection of preimplantation embryos were not successful. In contrast, infection of bovine oocytes with lentiviral vectors carrying an enhanced green fluorescent protein (eGFP) expression cassette followed by in vitro fertilization resulted in the birth of transgenic calves. Furthermore, all of the calves generated by infection of oocytes were transgenic, and 100% of these animals expressed eGFP as detected by in vivo imaging and Western blotting. In addition, a transgenic calf was produced by infection of fetal fibroblasts followed by nuclear transfer into enucleated oocytes. Taken together, after adjusting lentiviral transgenesis to cattle, unprecedented high transgenesis and expression rates were achieved.
Hofmann
Andreas
A
Department of Pharmacy, Institute for Pharmacology, Center for Drug Research, Ludwig-Maximilians University, Butenandstrasse 5(C), 81377 Munich, Germany.
Zakhartchenko
Valeri
V
Weppert
Myriam
M
Sebald
Heidi
H
Wenigerkind
Hendrik
H
Brem
Gottfried
G
Wolf
Eckhard
E
Pfeifer
Alexander
A
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D002417:Cattle; D005260:Female; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D009865:Oocytes / Q000502:physiology*; D011247:Pregnancy; D027724:Reproductive Techniques, Assisted / Q000662:veterinary*; D019076:Transgenes
D049452:Green Fluorescent Proteins