Efficient production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation
- Marialuisa Lavitrano*,†,
- Maria Laura Bacci‡,
- Monica Forni‡,
- Davide Lazzereschi*,
- Carla Di Stefano*,§,
- Daniela Fioretti*,§,
- Paola Giancotti*,§,
- Gabriella Marfé*,§,
- Loredana Pucci*,§,
- Luigina Renzi*,§,
- Hongjun Wang*,§,
- Antonella Stoppacciaro*,
- Giorgio Stassi¶,
- Massimo Sargiacomo∥,
- Paola Sinibaldi**,
- Valeria Turchi*,
- Roberto Giovannoni††,
- Giacinto Della Casa‡‡,
- Eraldo Seren‡, and
- Giancarlo Rossi‡‡
- *Dipartimento di Medicina Sperimentale, Università “La Sapienza,” 00161 Roma, Italy; ‡Dipartimento di Morfofisiologia Veterinaria e Produzioni Animali, Università di Bologna, 40064 Bologna, Italy; ¶Dipartimento di Discipline Chirurgiche, Anatomiche e Oncologiche, Università di Palermo, 90127 Palermo, Italy; ∥Istituto Superiore di Sanità, 00161 Roma, Italy; **Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Roma “Tor Vergata,” 00133 Roma, Italy; ††Dipartimento di Medicina Sperimentale, Ambientale e Biotecnologie Mediche, Università di Milano-Bicocca, 20052 Milano, Italy; and‡‡ Istituto Sperimentale per la Zootecnia, 00016 Monterotondo, Italy
-
Communicated by John C. Gerhart, University of California, Berkeley, CA (received for review August 3, 2002)
Abstract
A large number of hDAF transgenic pigs to be used for xenotransplantation research were generated by using sperm-mediated gene transfer (SMGT). The efficiency of transgenesis obtained with SMGT was much greater than with any other method. In the experiments reported, up to 80% of pigs had the transgene integrated into the genome. Most of the pigs carrying the hDAF gene transcribed it in a stable manner (64%). The great majority of pigs that transcribed the gene expressed the protein (83%). The hDAF gene was transmitted to progeny. Expression was stable and found in caveolae as it is in human cells. The expressed gene was functional based on in vitro experiments performed on peripheral blood mononuclear cells. These results show that our SMGT approach to transgenesis provides an efficient procedure for studies involving large animal models.
Footnotes
-
↵ † To whom correspondence should be sent at the present address: Dipartimento di Medicina Sperimentale, Ambientale e Biotecnologie Mediche, Università di Milano-Bicocca, Via Cadore 48, 20052 Milano, Italy. E-mail: marialuisa.lavitrano{at}unimib.it.
-
↵ § C.D.S., D.F., P.G., G.M., L.P., L.R., and H.W. contributed equally to this work.
- Abbreviations:
-
SMGT, sperm-mediated gene transfer
-
RT, room temperature
-
hDAF, human decay accelerating factor
-
FISH, fluorescence in situ hybridization
-
- Copyright © 2002, The National Academy of Sciences
