Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 19 December 1997:
Vol. 278. no. 5346, pp. 2130 - 2133
DOI: 10.1126/science.278.5346.2130
Prev | Table of Contents | Next

Reports

Human Factor IX Transgenic Sheep Produced by Transfer of Nuclei from Transfected Fetal Fibroblasts

Angelika E. Schnieke, * Alexander J. Kind, William A. Ritchie, Karen Mycock, Angela R. Scott, Marjorie Ritchie, Ian Wilmut, Alan Colman, Keith H. S. Campbell dagger

Ovine primary fetal fibroblasts were cotransfected with a neomycin resistance marker gene (neo) and a human coagulation factor IX genomic construct designed for expression of the encoded protein in sheep milk. Two cloned transfectants and a population of neomycin (G418)-resistant cells were used as donors for nuclear transfer to enucleated oocytes. Six transgenic lambs were liveborn: Three produced from cloned cells contained factor IX and neo transgenes, whereas three produced from the uncloned population contained the marker gene only. Somatic cells can therefore be subjected to genetic manipulation in vitro and produce viable animals by nuclear transfer. Production of transgenic sheep by nuclear transfer requires fewer than half the animals needed for pronuclear microinjection.

A. E. Schnieke, A. J. Kind, K. Mycock, A. R. Scott, A. Colman, PPL Therapeutics, Roslin, Midlothian, EH25 9PP, Scotland, UK.
W. A. Ritchie, M. Ritchie, I. Wilmut, K. H. S. Campbell, Roslin Institute, Roslin, Midlothian, EH25 9PS, Scotland, UK.
*   To whom correspondence should be addressed.

dagger    Present address: PPL Therapeutics, Roslin, Midlothian, EH25 9PP, Scotland, UK.

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
The endometrium responds differently to cloned versus fertilized embryos.
S. Bauersachs, S. E. Ulbrich, V. Zakhartchenko, M. Minten, M. Reichenbach, H.-D. Reichenbach, H. Blum, T. E. Spencer, and E. Wolf (2009)
PNAS 106, 5681-5686
   Abstract »    Full Text »    PDF »
Nuclear Transfer-Mediated Rescue of the Nuclear Genome of Nonviable Mouse Cells Frozen Without Cryoprotectant.
J. Li and P. Mombaerts (2008)
Biol Reprod 79, 588-593
   Abstract »    Full Text »    PDF »
Rabbit somatic cell cloning: effects of donor cell type, histone acetylation status and chimeric embryo complementation.
F. Yang, R. Hao, B. Kessler, G. Brem, E. Wolf, and V. Zakhartchenko (2007)
Reproduction 133, 219-230
   Abstract »    Full Text »    PDF »
Rabbits generated from fibroblasts through nuclear transfer..
S. Li, X. Chen, Z. Fang, J. Shi, and H. Z Sheng (2006)
Reproduction 131, 1085-1090
   Abstract »    Full Text »    PDF »
Functional disruption of the prion protein gene in cloned goats..
G. Yu, J. Chen, H. Yu, S. Liu, J. Chen, X. Xu, H. Sha, X. Zhang, G. Wu, S. Xu, et al. (2006)
J. Gen. Virol. 87, 1019-1027
   Abstract »    Full Text »    PDF »
Apoptosis in rabbit embryos produced by fertilization or nuclear transfer with fibroblasts and cumulus cells.
S.-Z. Liu, L.-J. Yao, M.-X. Jiang, Z.-L. Lei, L.-S. Zhang, Y.-L. Zhang, Q.-Y. Sun, Y.-L. Zheng, X.-F. Song, and D.-Y. Chen (2005)
Reproduction 130, 359-366
   Abstract »    Full Text »    PDF »
Transgenic Mice Produced by Retroviral Transduction of Male Germ Line Stem Cells In Vivo.
M. Kanatsu-Shinohara, S. Toyokuni, and T. Shinohara (2004)
Biol Reprod 71, 1202-1207
   Abstract »    Full Text »    PDF »
The first cell cycle after transfer of somatic cell nuclei in a non-human primate.
S.-C. Ng, N. Chen, W.-Y. Yip, S.-L. Liow, G.-Q. Tong, B. Martelli, L. G. Tan, and P. Martelli (2004)
Development 131, 2475-2484
   Abstract »    Full Text »    PDF »
Cloned transgenic farm animals produce a bispecific antibody for T cell-mediated tumor cell killing.
L. Grosse-Hovest, S. Muller, R. Minoia, E. Wolf, V. Zakhartchenko, H. Wenigerkind, C. Lassnig, U. Besenfelder, M. Muller, S. D. Lytton, et al. (2004)
PNAS 101, 6858-6863
   Abstract »    Full Text »    PDF »
Production of Nuclear Transfer-Derived Piglets Using Porcine Fetal Fibroblasts Transfected with the Enhanced Green Fluorescent Protein.
S. Hyun, G. Lee, D. Kim, H. Kim, S. Lee, D. Nam, Y. Jeong, S. Kim, S. Yeom, S. Kang, et al. (2003)
Biol Reprod 69, 1060-1068
   Abstract »    Full Text »    PDF »
Transgene Expression of Green Fluorescent Protein and Germ Line Transmission in Cloned Calves Derived from In Vitro-Transfected Somatic Cells.
V. Bordignon, R. Keyston, A. Lazaris, A. S. Bilodeau, J. H.F. Pontes, D. Arnold, G. Fecteau, C. Keefer, and L. C. Smith (2003)
Biol Reprod 68, 2013-2023
   Abstract »    Full Text »    PDF »
Mammary expression of new genes to combat mastitis.
D. E. Kerr and O. Wellnitz (2003)
J Anim Sci 81, 38-47
   Abstract »    Full Text »    PDF »
Effect of Nutrition of Oocyte Donor on the Outcomes of Somatic Cell Nuclear Transfer in the Sheep.
T. T. Peura, D. O. Kleemann, S. R. Rudiger, G. S. Nattrass, C. J. McLaughlan, and S. K. Walker (2003)
Biol Reprod 68, 45-50
   Abstract »    Full Text »    PDF »
Efficient Production of Transgenic Cloned Calves Using Preimplantation Screening.
S.-H. Chen, T. D. Vaught, J. A. Monahan, J. Boone, E. Emslie, P. M. Jobst, A. E. Lamborn, A. Schnieke, L. Robertson, A. Colman, et al. (2002)
Biol Reprod 67, 1488-1492
   Abstract »    Full Text »    PDF »
Aberrant Allocations of Inner Cell Mass and Trophectoderm Cells in Bovine Nuclear Transfer Blastocysts.
D.-B. Koo, Y.-K. Kang, Y.-H. Choi, J. S. Park, H.-N. Kim, K. B. Oh, D.-S. Son, H. Park, K.-K. Lee, and Y.-M. Han (2002)
Biol Reprod 67, 487-492
   Abstract »    Full Text »    PDF »
Fate of Donor Mitochondrial DNA in Cloned Bovine Embryos Produced by Microinjection of Cumulus Cells.
J. T. Do, J. W. Lee, B. Y. Lee, S. B. Kim, Z. Y. Ryoo, H. T. Lee, and K. S. Chung (2002)
Biol Reprod 67, 555-560
   Abstract »    Full Text »    PDF »
Nuclei of Nonviable Ovine Somatic Cells Develop into Lambs after Nuclear Transplantation.
P. Loi, M. Clinton, B. Barboni, J. Fulka Jr., P. Cappai, R. Feil, R. M. Moor, and G. Ptak (2002)
Biol Reprod 67, 126-132
   Abstract »    Full Text »    PDF »
In Vitro Development of Bovine Nuclear Transfer Embryos from Transgenic Clonal Lines of Adult and Fetal Fibroblast Cells of the Same Genotype.
S. Arat, J. Gibbons, S. J. Rzucidlo, D. S. Respess, M. Tumlin, and S. L. Stice (2002)
Biol Reprod 66, 1768-1774
   Abstract »    Full Text »    PDF »
Reproductive Characteristics of Cloned Heifers Derived from Adult Somatic Cells.
B.P. Enright, M. Taneja, D. Schreiber, J. Riesen, X.C. Tian, J.E. Fortune, and X. Yang (2002)
Biol Reprod 66, 291-296
   Abstract »    Full Text »    PDF »
Cloned Transgenic Offspring Resulting from Somatic Cell Nuclear Transfer in the Goat: Oocytes Derived from Both Follicle-Stimulating Hormone-Stimulated and Nonstimulated Abattoir-Derived Ovaries.
B. C. Reggio, A. N. James, H. L. Green, W. G. Gavin, E. Behboodi, Y. Echelard, and R. A. Godke (2001)
Biol Reprod 65, 1528-1533
   Abstract »    Full Text »    PDF »
Typical Demethylation Events in Cloned Pig Embryos. CLUES ON SPECIES-SPECIFIC DIFFERENCES IN EPIGENETIC REPROGRAMMING OF A CLONED DONOR GENOME.
Y.-K. Kang, D.-B. Koo, J. S. Park, Y.-H. Choi, H.-N. Kim, W.-K. Chang, K.-K. Lee, and Y.-M. Han (2001)
J. Biol. Chem. 276, 39980-39984
   Abstract »    Full Text »    PDF »
Nuclear Cloning and Epigenetic Reprogramming of the Genome.
W. M. Rideout III, K. Eggan, and R. Jaenisch (2001)
Science 293, 1093-1098
   Abstract »    Full Text »    PDF »
Clonal Lines of Transgenic Fibroblast Cells Derived from the Same Fetus Result in Different Development When Used for Nuclear Transfer in Pigs.
B. Kühholzer, R.J. Hawley, L. Lai, D. Kolber-Simonds, and R.S. Prather (2001)
Biol Reprod 64, 1695-1698
   Abstract »    Full Text »
Effect of Fibroblast Donor Cell Age and Cell Cycle on Development of Bovine Nuclear Transfer Embryos In Vitro.
P. Kasinathan, J. G. Knott, P. N. Moreira, A. S. Burnside, D. Joseph Jerry, and J. M. Robl (2001)
Biol Reprod 64, 1487-1493
   Abstract »    Full Text »
Generation of Dwarf Goat (Capra hircus) Clones Following Nuclear Transfer with Transfected and Nontransfected Fetal Fibroblasts and In Vitro-Matured Oocytes.
C.L. Keefer, H. Baldassarre, R. Keyston, B. Wang, B. Bhatia, A.S. Bilodeau, J.F. Zhou, M. Leduc, B.R. Downey, A. Lazaris, et al. (2001)
Biol Reprod 64, 849-856
   Abstract »    Full Text »
Culture of Preimplantation Mouse Embryos Affects Fetal Development and the Expression of Imprinted Genes.
S. Khosla, W. Dean, D. Brown, W. Reik, and R. Feil (2001)
Biol Reprod 64, 918-926
   Abstract »    Full Text »
Fertile and diploid nuclear transplants derived from embryonic cells of a small laboratory fish, medaka (Oryziaslatipes).
Y. Wakamatsu, B. Ju, I. Pristyaznhyuk, K. Niwa, T. Ladygina, M. Kinoshita, K. Araki, and K. Ozato (2001)
PNAS 98, 1071-1076
   Abstract »    Full Text »    PDF »
Cloned Mice from Fetal Fibroblast Cells Arrested at Metaphase by a Serial Nuclear Transfer.
Y. Ono, N. Shimozawa, M. Ito, and T. Kono (2001)
Biol Reprod 64, 44-50
   Abstract »    Full Text »
Development of Cloned Embryos from Adult Rabbit Fibroblasts: Effect of Activation Treatment and Donor Cell Preparation.
A. Dinnyés, Y. Dai, M. Barber, L. Liu, J. Xu, P. Zhou, and X. Yang (2001)
Biol Reprod 64, 257-263
   Abstract »    Full Text »
Evidence for Placental Abnormality as the Major Cause of Mortality in First-Trimester Somatic Cell Cloned Bovine Fetuses.
J. R. Hill, R. C. Burghardt, K. Jones, C. R. Long, C. R. Looney, T. Shin, T. E. Spencer, J. A. Thompson, Q. A. Winger, and M. E. Westhusin (2000)
Biol Reprod 63, 1787-1794
   Abstract »    Full Text »
In Vitro Development of Reconstructed Porcine Oocytes after Somatic Cell Nuclear Transfer.
D.-B. Koo, Y.-K. Kang, Y.-H. Choi, J. S. Park, S.-K. Han, I. Y. Park, S.-U. Kim, K.-K. Lee, D.-S. Son, W.-K. Chang, et al. (2000)
Biol Reprod 63, 986-992
   Abstract »    Full Text »
Advances in Livestock Nuclear Transfer.
B. Kühholzer and R. S. Prather (2000)
Experimental Biology and Medicine 224, 240-245
   Abstract »    Full Text »
Pig Cloning by Microinjection of Fetal Fibroblast Nuclei.
A. Onishi, M. Iwamoto, T. Akita, S. Mikawa, K. Takeda, T. Awata, H. Hanada, and A. C. F. Perry (2000)
Science 289, 1188-1190
   Abstract »    Full Text »
Production of Male Cloned Mice from Fresh, Cultured, and Cryopreserved Immature Sertoli Cells.
A. Ogura, K. Inoue, N. Ogonuki, A. Noguchi, K. Takano, R. Nagano, O. Suzuki, J. Lee, F. Ishino, and J. Matsuda (2000)
Biol Reprod 62, 1579-1584
   Abstract »    Full Text »
Establishment of a Porcine Cell Line from In Vitro-Produced Blastocysts and Transfer of the Cells into Enucleated Oocytes.
K. Miyoshi, Y. Taguchi, Y. Sendai, H. Hoshi, and E. Sato (2000)
Biol Reprod 62, 1640-1646
   Abstract »    Full Text »
Development Rates of Male Bovine Nuclear Transfer Embryos Derived from Adult and Fetal Cells.
J. R. Hill, Q. A. Winger, C. R. Long, C. R. Looney, J. A. Thompson, and M. E. Westhusin (2000)
Biol Reprod 62, 1135-1140
   Abstract »    Full Text »
Pre-selection of integration sites imparts repeatable transgene expression.
H. Wallace, R. Ansell, J. Clark, and J. McWhir (2000)
Nucleic Acids Res. 28, 1455-1464
   Abstract »    Full Text »    PDF »
From the Cover: Six cloned calves produced from adult fibroblast cells after long-term culture.
C. Kubota, H. Yamakuchi, J. Todoroki, K. Mizoshita, N. Tabara, M. Barber, and X. Yang (2000)
PNAS 97, 990-995
   Abstract »    Full Text »    PDF »
Cell Cycle Synchronization of Porcine Fetal Fibroblasts: Effects of Serum Deprivation and Reversible Cell Cycle Inhibitors.
W.A. Kues, M. Anger, J.W. Carnwath, D. Paul, J. Motlik, and H. Niemann (2000)
Biol Reprod 62, 412-419
   Abstract »    Full Text »
Double trans-chromosomic mice: Maintenance of two individual human chromosome fragments containing Ig heavy and kappa loci and expression of fully human antibodies.
K. Tomizuka, T. Shinohara, H. Yoshida, H. Uejima, A. Ohguma, S. Tanaka, K. Sato, M. Oshimura, and I. Ishida (2000)
PNAS 97, 722-727
   Abstract »    Full Text »    PDF »
Genetic Analysis of Inherited Hypertension in the Rat.
J. P. Rapp (2000)
Physiol Rev 80, 135-172
   Abstract »    Full Text »    PDF »
Mice cloned from embryonic stem cells.
T. Wakayama, I. Rodriguez, A. C. F. Perry, R. Yanagimachi, and P. Mombaerts (1999)
PNAS 96, 14984-14989
   Abstract »    Full Text »    PDF »
Bovine Oocyte Cytoplasm Supports Development of Embryos Produced by Nuclear Transfer of Somatic Cell Nuclei from Various Mammalian Species.
T. Dominko, M. Mitalipova, B. Haley, Z. Beyhan, E. Memili, B. McKusick, and N. L. First (1999)
Biol Reprod 60, 1496-1502
   Abstract »    Full Text »
Production of Cloned Calves Following Nuclear Transfer with Cultured Adult Mural Granulosa Cells.
D. N. Wells, P. M. Misica, and H. R. Tervit (1999)
Biol Reprod 60, 996-1005
   Abstract »    Full Text »
Nuclear Transfer in the Rhesus Monkey: Practical and Basic Implications.
D. P. Wolf, L. Meng, N. Ouhibi, and M. Zelinski-Wooten (1999)
Biol Reprod 60, 199-204
   Abstract »    Full Text »    PDF »
Cryopreservation of Bovine Pre-Morula-Stage In Vitro Matured/In Vitro Fertilized Embryos after Delipidation and before Use in Nucleus Transfer.
H. Ushijima, H. Yamakawa, and H. Nagashima (1999)
Biol Reprod 60, 534-539
   Abstract »    Full Text »    PDF »
Eight Calves Cloned from Somatic Cells of a Single Adult.
Y. Kato, T. Tani, Y. Sotomaru, K. Kurokawa, J. Kato, H. Doguchi, H. Yasue, and Y. Tsunoda (1998)
Science 282, 2095-2098
   Abstract »    Full Text »
Altered imprinted gene methylation and expression in completely ES cell-derived mouse fetuses: association with aberrant phenotypes.
W Dean, L Bowden, A Aitchison, J Klose, T Moore, J. Meneses, W Reik, and R Feil (1998)
Development 125, 2273-2282
   Abstract »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)