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.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Science 14 May 1999:
Vol. 284. no. 5417, pp. 1180 - 1183
DOI: 10.1126/science.284.5417.1180

Reports

Mammalian Transgenesis by Intracytoplasmic Sperm Injection

Anthony C. F. Perry, 1* Teruhiko Wakayama, 1 Hidefumi Kishikawa, 1 Tsuyoshi Kasai, 1 Masaru Okabe, 2 Yutaka Toyoda, 3 Ryuzo Yanagimachi 1

Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or beta -galactosidase reporter produced 64 to 94 percent transgene-expressing embryos, reflecting DNA-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos in the GFP series generated about 20 percent offspring expressing the integrated transgene. These data indicate that exogenous DNA can reproducibly be delivered into an oocyte by microinjected spermatozoa and suggest an adaptable method of transgenesis.

1 Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu, HI 96822, USA.
2 Genome Information Research Center, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871, Japan.
3 The Research Center for Protozoan Molecular Immunology, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080, Japan.
*   To whom correspondence should be addressed. E-mail: perry{at}hawaii.edu


Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Assessment of chromosomal integrity using a novel live-cell imaging technique in mouse embryos produced by intracytoplasmic sperm injection.
K. Yamagata, R. Suetsugu, and T. Wakayama (2009)
Hum. Reprod.
   Abstract »    Full Text »    PDF »
Production of normal mice from spermatozoa denatured with high alkali treatment before ICSI.
C. Li, E. Mizutani, T. Ono, and T. Wakayama (2009)
Reproduction 137, 779-792
   Abstract »    Full Text »    PDF »
Effects of disulfide bond reducing agents on sperm chromatin structural integrity and developmental competence of in vitro matured oocytes after intracytoplasmic sperm injection in pigs.
W.-M. Cheng, L. An, Z.-H. Wu, Y.-B. Zhu, J.-H. Liu, H.-M. Gao, X.-H. Li, S.-J. Zheng, D.-B. Chen, and J.-H. Tian (2009)
Reproduction 137, 633-643
   Abstract »    Full Text »    PDF »
Germ Cell Research: A Personal Perspective.
R. Yanagimachi (2009)
Biol Reprod 80, 204-218
   Abstract »    Full Text »    PDF »
Increasing the Cell Number of Host Tetraploid Embryos Can Improve the Production of Mice Derived from Embryonic Stem Cells.
H. Ohta, Y. Sakaide, K. Yamagata, and T. Wakayama (2008)
Biol Reprod 79, 486-492
   Abstract »    Full Text »    PDF »
Sperm-mediated 'reverse' gene transfer: a role of reverse transcriptase in the generation of new genetic information.
C. Spadafora (2008)
Hum. Reprod. 23, 735-740
   Abstract »    Full Text »    PDF »
Broad, ectopic expression of the sperm protein PLCZ1 induces parthenogenesis and ovarian tumours in mice.
N. Yoshida, M. Amanai, T. Fukui, E. Kajikawa, M. Brahmajosyula, A. Iwahori, Y. Nakano, S. Shoji, J. Diebold, H. Hessel, et al. (2007)
Development 134, 3941-3952
   Abstract »    Full Text »    PDF »
Genomic DNA Damage in Mouse Transgenesis.
Y. Yamauchi, B. Doe, A. Ajduk, and M. A Ward (2007)
Biol Reprod 77, 803-812
   Abstract »    Full Text »    PDF »
Lentiviral vectors: are they the future of animal transgenesis?.
F. Park (2007)
Physiol Genomics 31, 159-173
   Abstract »    Full Text »    PDF »
Altered protamine expression and diminished spermatogenesis: what is the link?.
D. T. Carrell, B. R. Emery, and S. Hammoud (2007)
Hum. Reprod. Update 13, 313-327
   Abstract »    Full Text »    PDF »
Effect of Transgene Concentration, Flanking Matrix Attachment Regions, and RecA-Coating on the Efficiency of Mouse Transgenesis Mediated by Intracytoplasmic Sperm Injection.
P. N. Moreira, M. Perez-Crespo, M. A. Ramirez, J. Pozueta, L. Montoliu, and A. Gutierrez-Adan (2007)
Biol Reprod 76, 336-343
   Abstract »    Full Text »    PDF »
Injection of Mammalian Metaphase II Oocytes with Short Interfering RNAs to Dissect Meiotic and Early Mitotic Events.
M. Amanai, S. Shoji, N. Yoshida, M. Brahmajosyula, and A. C.F. Perry (2006)
Biol Reprod 75, 891-898
   Abstract »    Full Text »    PDF »
A Restricted Role for Sperm-Borne MicroRNAs in Mammalian Fertilization.
M. Amanai, M. Brahmajosyula, and A. C.F. Perry (2006)
Biol Reprod 75, 877-884
   Abstract »    Full Text »    PDF »
Production of germline transgenic chickens expressing enhanced green fluorescent protein using a MoMLV-based retrovirus vector.
B. C. Koo, M. S. Kwon, B. R. Choi, J.-H. Kim, S.-K. Cho, S. H. Sohn, E. J. Cho, H. T. Lee, W. Chang, I. Jeon, et al. (2006)
FASEB J 20, 2251-2260
   Abstract »    Full Text »    PDF »
DNase I activity in pig MII oocytes: implications in transgenesis..
A. Zannoni, M. Spinaci, C. Bernardini, M. L. Bacci, E. Seren, M. Mattioli, and M. Forni (2006)
Reproduction 131, 461-468
   Abstract »    Full Text »    PDF »
Morphologic changes in boar sperm nuclei with reduced disulfide bonds in electrostimulated porcine oocytes..
M. Nakai, N. Kashiwazaki, A. Takizawa, N. Maedomari, M. Ozawa, J. Noguchi, H. Kaneko, M. Shino, and K. Kikuchi (2006)
Reproduction 131, 603-611
   Abstract »    Full Text »    PDF »
Inadvertent transgenesis by conventional ICSI in mice.
P. N. Moreira, R. Fernandez-Gonzalez, D. Rizos, M. Ramirez, M. Perez-Crespo, and A. Gutierrez-Adan (2005)
Hum. Reprod. 20, 3313-3317
   Abstract »    Full Text »    PDF »
Tn5 Transposase-Mediated Mouse Transgenesis.
R. Suganuma, P. Pelczar, J. F. Spetz, B. Hohn, R. Yanagimachi, and S. Moisyadi (2005)
Biol Reprod 73, 1157-1163
   Abstract »    Full Text »    PDF »
Wild-Type Measles Virus Infection in Human CD46/CD150-Transgenic Mice: CD11c-Positive Dendritic Cells Establish Systemic Viral Infection.
M. Shingai, N. Inoue, T. Okuno, M. Okabe, T. Akazawa, Y. Miyamoto, M. Ayata, K. Honda, M. Kurita-Taniguchi, M. Matsumoto, et al. (2005)
J. Immunol. 175, 3252-3261
   Abstract »    Full Text »    PDF »
Developmental Regulation and In Vitro Culture Effects on Expression of DNA Repair and Cell Cycle Checkpoint Control Genes in Rhesus Monkey Oocytes and Embryos.
P. Zheng, R. D. Schramm, and K. E. Latham (2005)
Biol Reprod 72, 1359-1369
   Abstract »    Full Text »    PDF »
Establishment of Male and Female Nuclear Transfer Embryonic Stem Cell Lines from Different Mouse Strains and Tissues.
S. Wakayama, H. Ohta, S. Kishigami, N. Van Thuan, T. Hikichi, E. Mizutani, M. Miyake, and T. Wakayama (2005)
Biol Reprod 72, 932-936
   Abstract »    Full Text »    PDF »
Efficient Generation of Transgenic Mice with Intact Yeast Artificial Chromosomes by Intracytoplasmic Sperm Injection.
P. N. Moreira, P. Giraldo, P. Cozar, J. Pozueta, A. Jimenez, L. Montoliu, and A. Gutierrez-Adan (2004)
Biol Reprod 71, 1943-1947
   Abstract »    Full Text »    PDF »
The effect of human immunodeficiency virus on sperm parameters and the outcome of intrauterine insemination following sperm washing.
J. D.M. Nicopoullos, P. A. Almeida, J. W.A. Ramsay, and C. Gilling-Smith (2004)
Hum. Reprod. 19, 2289-2297
   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 »
Spermatogenesis and Germ Cell Transgene Expression in Xenografted Bovine Testicular Tissue.
J. M. Oatley, D. M. de Avila, J. J. Reeves, and D. J. McLean (2004)
Biol Reprod 71, 494-501
   Abstract »    Full Text »    PDF »
Similar Time Restriction for Intracytoplasmic Sperm Injection and Round Spermatid Injection into Activated Oocytes for Efficient Offspring Production.
S. Kishigami, S. Wakayama, N. Van Thuan, and T. Wakayama (2004)
Biol Reprod 70, 1863-1869
   Abstract »    Full Text »    PDF »
Transgenic zebrafish produced by retroviral infection of in vitro-cultured sperm.
K. Kurita, S. M. Burgess, and N. Sakai (2004)
PNAS 101, 1263-1267
   Abstract »    Full Text »    PDF »
Children conceived using ICSI do not have an increased risk of delayed mental development at 5 years of age.
G.I. Leslie, F.L. Gibson, C. McMahon, J. Cohen, D.M. Saunders, and C. Tennant (2003)
Hum. Reprod. 18, 2067-2072
   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 »
Somatic Histones Are Components of the Perinuclear Theca in Bovine Spermatozoa.
P. R. Tovich and R. J. Oko (2003)
J. Biol. Chem. 278, 32431-32438
   Abstract »    Full Text »    PDF »
Expression of Foreign DNA Is Associated with Paternal Chromosome Degradation in Intracytoplasmic Sperm Injection-Mediated Transgenesis in the Mouse.
M. A. Szczygiel, S. Moisyadi, and W. S. Ward (2003)
Biol Reprod 68, 1903-1910
   Abstract »    Full Text »    PDF »
The Making of "Transgenic Spermatozoa".
C. Celebi, T. Guillaudeux, P. Auvray, V. Vallet-Erdtmann, and B. Jegou (2003)
Biol Reprod 68, 1477-1483
   Abstract »    Full Text »    PDF »
Viable Piglets Generated from Porcine Oocytes Matured In Vitro and Fertilized by Intracytoplasmic Sperm Head Injection.
M. Nakai, N. Kashiwazaski, A. Takizawa, Y. Hayashi, E. Nakatsukasa, D.-i. Fuchimoto, J. Noguchi, H. Kaneko, M. Shino, and K. Kikuchi (2003)
Biol Reprod 68, 1003-1008
   Abstract »    Full Text »    PDF »
Efficient production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation.
M. Lavitrano, M. L. Bacci, M. Forni, D. Lazzereschi, C. Di Stefano, D. Fioretti, P. Giancotti, G. Marfe, L. Pucci, L. Renzi, et al. (2002)
PNAS 99, 14230-14235
   Abstract »    Full Text »    PDF »
Prenatal testing in ICSI pregnancies: incidence of chromosomal anomalies in 1586 karyotypes and relation to sperm parameters.
M. Bonduelle, E. Van Assche, H. Joris, K. Keymolen, P. Devroey, A. Van Steirteghem, and I. Liebaers (2002)
Hum. Reprod. 17, 2600-2614
   Abstract »    Full Text »    PDF »
Bovine Blastocyst Development from Oocytes Injected with Freeze-Dried Spermatozoa.
L. Keskintepe, G. Pacholczyk, A. Machnicka, K. Norris, M. A. Curuk, I. Khan, and B. G. Brackett (2002)
Biol Reprod 67, 409-415
   Abstract »    Full Text »    PDF »
Mosaic Gene Expression in Nuclear Transfer-Derived Embryos and the Production of Cloned Transgenic Pigs from Ear-Derived Fibroblasts.
K.-W. Park, L. Lai, H.-T. Cheong, R. Cabot, Q.-Y. Sun, G. Wu, E. B. Rucker, D. Durtschi, A. Bonk, M. Samuel, et al. (2002)
Biol Reprod 66, 1001-1005
   Abstract »    Full Text »    PDF »
Effect of Partial Incision of the Zona Pellucida by Piezo-Micromanipulator for In Vitro Fertilization Using Frozen-Thawed Mouse Spermatozoa on the Developmental Rate of Embryos Transferred at the 2-Cell Stage.
Y. Kawase, T. Iwata, O. Ueda, N. Kamada, T. Tachibe, Y. Aoki, K.-i. Jishage, and H. Suzuki (2002)
Biol Reprod 66, 381-385
   Abstract »    Full Text »    PDF »
Should ICSI be the treatment of choice for all cases of in-vitro conception?: Considerations of fertilization and embryo development, cost ffectiveness and safety.
B. Ola, M. Afnan, K. Sharif, S. Papaioannou, N. Hammadieh, and C. L.R.Barratt (2001)
Hum. Reprod. 16, 2485-2490
   Abstract »    Full Text »    PDF »
Transgenic mice produced by retroviral transduction of male germ-line stem cells.
M. Nagano, C. J. Brinster, K. E. Orwig, B.-Y. Ryu, M. R. Avarbock, and R. L. Brinster (2001)
PNAS
   Abstract »    Full Text »    PDF »
Experimental inoculation of male mice with murine cytomegalovirus and effect on offspring.
L. Tebourbi, A.M. Courtot, R. Duchateau, A. Loeuillet, J. Testart, and I. Cerutti (2001)
Hum. Reprod. 16, 2041-2049
   Abstract »    Full Text »    PDF »
Fertilization of Eggs of Zebrafish, Danio rerio, by Intracytoplasmic Sperm Injection.
G. A. Poleo, R. S. Denniston, B. C. Reggio, R. A. Godke, and T. R. Tiersch (2001)
Biol Reprod 65, 961-966
   Abstract »    Full Text »    PDF »
ICSI choreography: fate of sperm structures after monospermic rhesus ICSI and first cell cycle implications.
J. Ramalho-Santos, P. Sutovsky, C. Simerly, R. Oko, G. M. Wessel, L. Hewitson, and G. Schatten (2000)
Hum. Reprod. 15, 2610-2620
   Abstract »    Full Text »    PDF »
Intracytoplasmic sperm injection and other aspects of new reproductive technologies.
A. G SUTCLIFFE (2000)
Arch. Dis. Child. 83, 98-101
   Full Text »    PDF »
Development of In Vivo-Matured Porcine Oocytes Following Intracytoplasmic Sperm Injection.
M. J. Martin (2000)
Biol Reprod 63, 109-112
   Abstract »    Full Text »
A simplified method of generating transgenic Xenopus.
D. B. Sparrow, B. Latinkic, and T. J. Mohun (2000)
Nucleic Acids Res. 28, e12
   Abstract »    Full Text »    PDF »
Foreign DNA transmission by ICSI: injection of spermatozoa bound with exogenous DNA results in embryonic GFP expression and live Rhesus monkey births.
A. W.S. Chan, C.M. Luetjens, T. Dominko, J. Ramalho-Santos, C. R. Simerly, L. Hewitson, and G. Schatten (2000)
Mol. Hum. Reprod. 6, 26-33
   Abstract »    Full Text »    PDF »
New Insights into Host Factor Requirements for Prokaryotic beta -Recombinase-mediated Reactions in Mammalian Cells.
V. Diaz, P. Servert, I. Prieto, M. A. Gonzalez, C. Martinez-A, J. C. Alonso, and A. Bernad (2001)
J. Biol. Chem. 276, 16257-16264
   Abstract »    Full Text »    PDF »
From the Cover: Transgenic mice produced by retroviral transduction of male germ-line stem cells.
M. Nagano, C. J. Brinster, K. E. Orwig, B.-Y. Ryu, M. R. Avarbock, and R. L. Brinster (2001)
PNAS 98, 13090-13095
   Abstract »    Full Text »    PDF »



To Advertise     Find Products


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