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Nucleic Acids Research, 1995, Vol. 23, No. 19 3816-3821
© 1995


MOLECULAR BIOLOGY

Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase

Yumi Kanegae1, Gwang Lee1, Yumi Sato1, Mieko Tanaka1, Michio Nakal1,3, Toshiyuki Sakaki3, Sumio Sugano2 and Izumu Saito1,*

1Laboaory of Molecular Genetics University of Tokyo 4-6-1 Shirokanedi, Minato-ku, Tokyo 108, Japan 2Deparment of Virology, institute of Medical Science, University of Tokyo 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan 3Discovery Research Laboratories III, Sumitomo Pharmaceuticals Research Center 3-1-98 Kasugade-naka, Konohana-ku, Osaka 554, Japan

*To whom correspondence should be addressed

Received August 8, 1995. Accepted August 23, 1995.

A recombinant adenovirus (Ad) expressing Cre recombinase derived from bacteriophage P1 was constructed. To assay the Cre activity in mammalian colls, another recombinant Ad bearing an on/off-switching reporter unit, where a LacZ-expression unit can be activated by the Cre-mediated exclsionai deletion of an interposed stuffer DNA, was also constructed. Co-infection experiments together with the Cre-expressing and the reporter recombinant Ads showed that the Cre-mediated switching of gene expression was detected in nearly 100% of cultured CV1, HeLa and Jurkat cells. These results suggest that the recombinant Ad efficiently expressed functional Cre and offers a basis for estabilshing a powerful on/off switching strategy of gene expression incultured mammalian cells and presumably in transgenic animals. The method is also applicable to construction of recombinant Ad bearing a gene the expression of which is deleterious to propagation of recombinant Ad.


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T. Kudo, S. Kure, K. Ikeda, A.-P. Xia, Y. Katori, M. Suzuki, K. Kojima, A. Ichinohe, Y. Suzuki, Y. Aoki, et al.
Transgenic expression of a dominant-negative connexin26 causes degeneration of the organ of Corti and non-syndromic deafness
Hum. Mol. Genet., May 1, 2003; 12(9): 995 - 1004.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
T. Ebihara, I. Kawabata, S. Usui, K. Sobue, and S. Okabe
Synchronized Formation and Remodeling of Postsynaptic Densities: Long-Term Visualization of Hippocampal Neurons Expressing Postsynaptic Density Proteins Tagged with Green Fluorescent Protein
J. Neurosci., March 15, 2003; 23(6): 2170 - 2181.
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J. Virol.Home page
T. Nakamura, K. Sato, and H. Hamada
Reduction of Natural Adenovirus Tropism to the Liver by both Ablation of Fiber-Coxsackievirus and Adenovirus Receptor Interaction and Use of Replaceable Short Fiber
J. Virol., February 15, 2003; 77(4): 2512 - 2521.
[Abstract] [Full Text] [PDF]


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HypertensionHome page
M. Yamasaki, J. Kawai, T. Nakaoka, T. Ogita, A. Tojo, and T. Fujita
Adrenomedullin Overexpression to Inhibit Cuff-Induced Arterial Intimal Formation
Hypertension, February 1, 2003; 41(2): 302 - 307.
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J. Virol.Home page
M. Tanaka, H. Kagawa, Y. Yamanashi, T. Sata, and Y. Kawaguchi
Construction of an Excisable Bacterial Artificial Chromosome Containing a Full-Length Infectious Clone of Herpes Simplex Virus Type 1: Viruses Reconstituted from the Clone Exhibit Wild-Type Properties In Vitro and In Vivo
J. Virol., December 20, 2002; 77(2): 1382 - 1391.
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Am. J. Physiol. Endocrinol. Metab.Home page
M. Tsuru, H. Katagiri, T. Asano, T. Yamada, S. Ohno, T. Ogihara, and Y. Oka
Role of PKC isoforms in glucose transport in 3T3-L1 adipocytes: insignificance of atypical PKC
Am J Physiol Endocrinol Metab, August 1, 2002; 283(2): E338 - E345.
[Abstract] [Full Text] [PDF]


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DiabetesHome page
Y. Tamori, J. Masugi, N. Nishino, and M. Kasuga
Role of Peroxisome Proliferator-Activated Receptor-{gamma} in Maintenance of the Characteristics of Mature 3T3-L1 Adipocytes
Diabetes, July 1, 2002; 51(7): 2045 - 2055.
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J. Biol. Chem.Home page
M. D. Johnson, X. Wu, N. Aithmitti, and R. S. Morrison
Peg3/Pw1 Is a Mediator between p53 and Bax in DNA Damage-induced Neuronal Death
J. Biol. Chem., June 14, 2002; 277(25): 23000 - 23007.
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J. Virol.Home page
Y. F. Jin, T. Ishibashi, A. Nomoto, and M. Masuda
Isolation and Analysis of Retroviral Integration Targets by Solo Long Terminal Repeat Inverse PCR
J. Virol., May 3, 2002; 76(11): 5540 - 5547.
[Abstract] [Full Text] [PDF]


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Hum Mol GenetHome page
S. Sato, M. Nakamura, D. H. Cho, S. J. Tapscott, H. Ozaki, and K. Kawakami
Identification of transcriptional targets for Six5: implication for the pathogenesis of myotonic dystrophy type 1
Hum. Mol. Genet., May 1, 2002; 11(9): 1045 - 1058.
[Abstract] [Full Text] [PDF]


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Br J OphthalmolHome page
M Akimoto, T Miyahara, J Arai, A Akimoto, H Hamada, Y Yoshida, and N Yoshimura
A new delivery system for 5-fluorouracil using prodrug and converting enzyme
Br J Ophthalmol, May 1, 2002; 86(5): 581 - 586.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
K. Katagiri, M. Hattori, N. Minato, and T. Kinashi
Rap1 Functions as a Key Regulator of T-Cell and Antigen-Presenting Cell Interactions and Modulates T-Cell Responses
Mol. Cell. Biol., February 15, 2002; 22(4): 1001 - 1015.
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J. Biol. Chem.Home page
H. Tang, Q. Hao, T. Fitzgerald, T. Sasaki, E. J. Landon, and T. Inagami
Pyk2/CAKbeta Tyrosine Kinase Activity-mediated Angiogenesis of Pulmonary Vascular Endothelial Cells
J. Biol. Chem., February 8, 2002; 277(7): 5441 - 5447.
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DiabetesHome page
N. Takahashi, T. Nemoto, R. Kimura, A. Tachikawa, A. Miwa, H. Okado, Y. Miyashita, M. Iino, T. Kadowaki, and H. Kasai
Two-Photon Excitation Imaging of Pancreatic Islets With Various Fluorescent Probes
Diabetes, February 1, 2002; 51(90001): S25 - 28.
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J. Neurosci.Home page
S. Okabe, T. Urushido, D. Konno, H. Okado, and K. Sobue
Rapid Redistribution of the Postsynaptic Density Protein PSD-Zip45 (Homer 1c) and Its Differential Regulation by NMDA Receptors and Calcium Channels
J. Neurosci., December 15, 2001; 21(24): 9561 - 9571.
[Abstract] [Full Text] [PDF]


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Int ImmunolHome page
M. Senoo, N. Mochida, L. Wang, Y. Matsumura, D. Suzuki, N. Takeda, Y. Shinkai, and S. Habu
Limited effect of chromatin remodeling on D{beta}-to-J{beta} recombination in CD4+CD8+ thymocyte: implications for a new aspect in the regulation of TCR {beta} gene recombination
Int. Immunol., November 1, 2001; 13(11): 1405 - 1414.
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J. Am. Soc. Nephrol.Home page
T. YOKOO, T. OHASHI, Y. UTSUNOMIYA, H. SHIBA, J. S. SHEN, Y. HISADA, Y. ETO, T. KAWAMURA, and T. HOSOYA
Inflamed Glomeruli--Specific Gene Activation that Uses Recombinant Adenovirus with the Cre/loxP System
J. Am. Soc. Nephrol., November 1, 2001; 12(11): 2330 - 2337.
[Abstract] [Full Text] [PDF]


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JCBHome page
Y. Liu, Z. Cseresnyes, W. R. Randall, and M. F. Schneider
Activity-dependent nuclear translocation and intranuclear distribution of NFATc in adult skeletal muscle fibers
J. Cell Biol., October 1, 2001; 155(1): 27 - 40.
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Cancer Res.Home page
S. Nishimura, M. Adachi, T. Ishida, T. Matsunaga, H. Uchida, H. Hamada, and K. Imai
Adenovirus-mediated Transfection of Caspase-8 Augments Anoikis and Inhibits Peritoneal Dissemination of Human Gastric Carcinoma Cells
Cancer Res., October 1, 2001; 61(19): 7009 - 7014.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
S. Okabe, A. Miwa, and H. Okado
Spine Formation and Correlated Assembly of Presynaptic and Postsynaptic Molecules
J. Neurosci., August 15, 2001; 21(16): 6105 - 6114.
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Cancer Res.Home page
K. Ueda, M. Iwahashi, M. Nakamori, M. Nakamura, I. Matsuura, H. Yamaue, and H. Tanimura
Carcinoembryonic Antigen-specific Suicide Gene Therapy of Cytosine Deaminase/5-Fluorocytosine Enhanced by the Cre/loxP System in the Orthotopic Gastric Carcinoma Model
Cancer Res., August 1, 2001; 61(16): 6158 - 6162.
[Abstract] [Full Text] [PDF]


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BloodHome page
T. Yokoo, T. Ohashi, Y. Utsunomiya, J. S. Shen, Y. Hisada, Y. Eto, T. Kawamura, and T. Hosoya
Genetically modified bone marrow continuously supplies anti-inflammatory cells and suppresses renal injury in mouse Goodpasture syndrome
Blood, July 1, 2001; 98(1): 57 - 64.
[Abstract] [Full Text] [PDF]


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Int ImmunolHome page
Y. Nonomura, H. Kohsaka, K. Nasu, Y. Terada, M.-a. Ikeda, and N. Miyasaka
Suppression of arthritis by forced expression of cyclin-dependent kinase inhibitor p21Cip1 gene into the joints
Int. Immunol., June 1, 2001; 13(6): 723 - 731.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Renal Physiol.Home page
S. Hanada, Y. Terada, S. Inoshita, S. Sasaki, S. M. Lohmann, A. Smolenski, and F. Marumo
Overexpression of protein kinase G using adenovirus inhibits cyclin E transcription and mesangial cell cycle
Am J Physiol Renal Physiol, May 1, 2001; 280(5): F851 - F859.
[Abstract] [Full Text] [PDF]


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Nucleic Acids ResHome page
M. Nakano, K. Odaka, M. Ishimura, S. Kondo, N. Tachikawa, J. Chiba, Y. Kanegae, and I. Saito
Efficient gene activation in cultured mammalian cells mediated by FLP recombinase-expressing recombinant adenovirus
Nucleic Acids Res., April 1, 2001; 29(7): e40 - e40.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
K. Kuwahara, Y. Saito, E. Ogawa, N. Takahashi, Y. Nakagawa, Y. Naruse, M. Harada, I. Hamanaka, T. Izumi, Y. Miyamoto, et al.
The Neuron-Restrictive Silencer Element-Neuron-Restrictive Silencer Factor System Regulates Basal and Endothelin 1-Inducible Atrial Natriuretic Peptide Gene Expression in Ventricular Myocytes
Mol. Cell. Biol., March 15, 2001; 21(6): 2085 - 2097.
[Abstract] [Full Text]


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Nucleic Acids ResHome page
T.-H. Cheng, C.-R. Chang, P. Joy, S. Yablok, and M. R. Gartenberg
Controlling gene expression in yeast by inducible site-specific recombination
Nucleic Acids Res., December 15, 2000; 28(24): e108 - e108.
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J. Immunol.Home page
K. Nasu, H. Kohsaka, Y. Nonomura, Y. Terada, H. Ito, K. Hirokawa, and N. Miyasaka
Adenoviral Transfer of Cyclin-Dependent Kinase Inhibitor Genes Suppresses Collagen-Induced Arthritis in Mice
J. Immunol., December 15, 2000; 165(12): 7246 - 7252.
[Abstract] [Full Text] [PDF]


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J. Virol.Home page
W. Satoh, Y. Hirai, K. Tamayose, and T. Shimada
Site-Specific Integration of an Adeno-Associated Virus Vector Plasmid Mediated by Regulated Expression of Rep Based on Cre-loxP Recombination
J. Virol., November 15, 2000; 74(22): 10631 - 10638.
[Abstract] [Full Text]


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J. Clin. Endocrinol. Metab.Home page
Y. Nagayama, H. Yokoi, K. Takeda, M. Hasegawa, E. Nishihara, H. Namba, S. Yamashita, and M. Niwa
Adenovirus-Mediated Tumor Suppressor p53 Gene Therapy for Anaplastic Thyroid Carcinoma in Vitro and in Vivo
J. Clin. Endocrinol. Metab., November 1, 2000; 85(11): 4081 - 4086.
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Arterioscler. Thromb. Vasc. Bio.Home page
H. Obara, A. Takayanagi, J. Hirahashi, K. Tanaka, G. Wakabayashi, K. Matsumoto, M. Shimazu, N. Shimizu, and M. Kitajima
Overexpression of Truncated I{kappa}B{alpha} Induces TNF-{alpha}-Dependent Apoptosis in Human Vascular Smooth Muscle Cells
Arterioscler Thromb Vasc Biol, October 1, 2000; 20(10): 2198 - 2204.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
K. Namikawa, M. Honma, K. Abe, M. Takeda, K. Mansur, T. Obata, A. Miwa, H. Okado, and H. Kiyama
Akt/Protein Kinase B Prevents Injury-Induced Motoneuron Death and Accelerates Axonal Regeneration
J. Neurosci., April 15, 2000; 20(8): 2875 - 2886.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
Y. Banzai, H. Miki, H. Yamaguchi, and T. Takenawa
Essential Role of Neural Wiskott-Aldrich Syndrome Protein in Neurite Extension in PC12 Cells and Rat Hippocampal Primary Culture Cells
J. Biol. Chem., April 14, 2000; 275(16): 11987 - 11992.
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J. Biol. Chem.Home page
M. Kawanishi, Y. Tamori, H. Okazawa, S. Araki, H. Shinoda, and M. Kasuga
Role of SNAP23 in Insulin-induced Translocation of GLUT4 in 3T3-L1 Adipocytes. MEDIATION OF COMPLEX FORMATION BETWEEN SYNTAXIN4 AND VAMP2
J. Biol. Chem., March 10, 2000; 275(11): 8240 - 8247.
[Abstract] [Full Text] [PDF]


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ScienceHome page
N. Kobayashi, T. Fujiwara, K. A. Westerman, Y. Inoue, M. Sakaguchi, H. Noguchi, M. Miyazaki, J. Cai, N. Tanaka, I. J. Fox, et al.
Prevention of Acute Liver Failure in Rats with Reversibly Immortalized Human Hepatocytes
Science, February 18, 2000; 287(5456): 1258 - 1262.
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EndocrinologyHome page
T. Nakanishi, T. Nishida, T. Shimo, K. Kobayashi, T. Kubo, T. Tamatani, K. Tezuka, and M. Takigawa
Effects of CTGF/Hcs24, a Product of a Hypertrophic Chondrocyte-Specific Gene, on the Proliferation and Differentiation of Chondrocytes in Culture
Endocrinology, January 1, 2000; 141(1): 264 - 273.
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J. Gen. Virol.Home page
Y. Shintani, H. Yotsuyanagi, K. Moriya, H. Fujie, T. Tsutsumi, Y. Kanegae, S. Kimura, I. Saito, and K. Koike
Induction of apoptosis after switch-on of the hepatitis B virus X gene mediated by the Cre/loxP recombination system
J. Gen. Virol., December 1, 1999; 80(12): 3257 - 3265.
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J. Biol. Chem.Home page
A. Asai, J.-h. Qiu, Y. Narita, S. Chi, N. Saito, N. Shinoura, H. Hamada, Y. Kuchino, and T. Kirino
High Level Calcineurin Activity Predisposes Neuronal Cells to Apoptosis
J. Biol. Chem., November 26, 1999; 274(48): 34450 - 34458.
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J. Biol. Chem.Home page
H. Kuwahara, N. Araki, K. Makino, N. Masuko, S. Honda, K. Kaibuchi, K. Fukunaga, E. Miyamoto, M. Ogawa, and H. Saya
A Novel NE-dlg/SAP102-associated Protein, p51-nedasin, Related to the Amidohydrolase Superfamily, Interferes with the Association between NE-dlg/SAP102 and N-Methyl-D-aspartate Receptor
J. Biol. Chem., November 5, 1999; 274(45): 32204 - 32214.
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