Efficient in vitro and in vivo excision of floxed sequences with a high-capacity adenoviral vector expressing Cre recombinase

被引:10
作者
Badorf, M
Edenhofer, F
Dries, V
Kochanek, S
Schiedner, G
机构
[1] Univ Cologne, Ctr Mol Med, ZMMK, D-50931 Cologne, Germany
[2] Univ Cologne, Inst Genet, D-50931 Cologne, Germany
[3] Univ Cologne, Inst Pathol, D-50931 Cologne, Germany
关键词
D O I
10.1002/gene.10099
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Conditional gene expression or gene disruption using Cre/loxP- or FLP/frt-based recombination systems are valuable tools for studying gene function in development and disease. Recombinant adenoviral vectors expressing Cre recombinase have been suggested as an alternative for deletion of floxed sequences. To further improve this approach we generated a high-capacity adenoviral (HC-Ad) vector expressing Cre (HC-Adcre). In this vector all viral coding. sequences are deleted resulting in decreased toxicity. In the present study HC-Adcre efficiently mediated recombination between two loxP sites located in the genome of a reporter cell line. When intravenously injected into ROSA26 reporter mice, a floxed sequence was excised in hepatocytes resulting in expression of the beta-gal reporter. Our data indicate that HC-Ad vectors expressing Cre effectively delete floxed sequences in vivo and have a significant potential as a tool for functional studies in mice. (C) 2002 Wiley-Liss, Inc.
引用
收藏
页码:119 / 124
页数:6
相关论文
共 30 条
[1]   Cre-mediated somatic site-specific recombination in mice [J].
Akagi, K ;
Sandig, V ;
Vooijs, M ;
VanderValk, M ;
Giovannini, M ;
Strauss, M ;
Berns, A .
NUCLEIC ACIDS RESEARCH, 1997, 25 (09) :1766-1773
[2]  
ANDERSON L, 1995, ADHES AGE, V38, P4
[3]  
[Anonymous], 1994, MANIPULATING MOUSE E
[4]   SITE-SPECIFIC RECOMBINATION MEDIATED BY AN ADENOVIRUS VECTOR EXPRESSING THE CRE RECOMBINASE PROTEIN - A MOLECULAR SWITCH FOR CONTROL OF GENE-EXPRESSION [J].
ANTON, M ;
GRAHAM, FL .
JOURNAL OF VIROLOGY, 1995, 69 (08) :4600-4606
[5]   Adenovirus-mediated regulable target gene expression in vivo [J].
Burcin, MM ;
Schiedner, G ;
Kochanek, S ;
Tsai, SY ;
O'Malley, BW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (02) :355-360
[6]   Liver-specific inactivation of the abetalipoproteinemia gene completely abrogates very low density lipoprotein low density lipoprotein production in a viable conditional knockout mouse [J].
Chang, BHJ ;
Liao, W ;
Li, L ;
Nakamuta, M ;
Mack, D ;
Chan, L .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (10) :6051-6055
[7]   AUTOMATED DNA SEQUENCING OF THE HUMAN HPRT LOCUS [J].
EDWARDS, A ;
VOSS, H ;
RICE, P ;
CIVITELLO, A ;
STEGEMANN, J ;
SCHWAGER, C ;
ZIMMERMANN, J ;
ERFLE, H ;
CASKEY, CT ;
ANSORGE, W .
GENOMICS, 1990, 6 (04) :593-608
[8]   EFFICIENT GENE ACTIVATION IN MAMMALIAN-CELLS BY USING RECOMBINANT ADENOVIRUS EXPRESSING SITE-SPECIFIC CRE RECOMBINASE [J].
KANEGAE, Y ;
LEE, G ;
SATO, Y ;
TANAKA, M ;
NAKAI, M ;
SAKAKI, T ;
SUGANO, S ;
SAITO, I .
NUCLEIC ACIDS RESEARCH, 1995, 23 (19) :3816-3821
[9]   Efficient gene activation system on mammalian cell chromosomes using recombinant adenovirus producing Cre recombinase [J].
Kanegae, Y ;
Takamori, K ;
Sato, Y ;
Lee, G ;
Nakai, M ;
Saito, I .
GENE, 1996, 181 (1-2) :207-212
[10]   Regulation of cre recombinase activity by the synthetic steroid RU 486 [J].
Kellendonk, C ;
Tronche, F ;
Monaghan, AP ;
Angrand, PO ;
Stewart, F ;
Schutz, G .
NUCLEIC ACIDS RESEARCH, 1996, 24 (08) :1404-1411