Efficient sequential gene regulation via FLP- and Cre-recombinase using adenovirus vector in mammalian cells including mouse ES cells

被引:11
作者
Kondo, Saki
Takahashi, Yuzuka
Shiozawa, Seiji
Ichise, Hirotake
Yoshida, Nobuaki
Kanegae, Yumi
Saito, Izumu
机构
[1] Univ Tokyo, Inst Med Sci, Genet Mol Lab, Minato Ku, Tokyo 1088639, Japan
[2] Univ Tokyo, Inst Med Sci, Lab Gene Express & Regulat, Minato Ku, Tokyo 1088639, Japan
关键词
gene regulation; Cre/loxP system; FLP/FRT system; adenovirus vector;
D O I
10.1111/j.1348-0421.2006.tb03850.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Site-specific recombinase is widely applied for the regulation of gene expression because its regulatory action is strict and efficient. However, each system can mediate regulation of only one gene at a time. Here, we demonstrate efficient "sequential" gene regulation using Cre- and FLP-expressing recombinant adenovirus (rAd) in two different monitor cell lines, for regulation of one gene (OFF-ON-OFF) and for two genes (ON-OFF and OFF-ON, independently). Generally, serial use of Cre- and FLP-expressing rAd tends to cause significant cytotoxicity, but we here described optimum dose of the rAds for serial regulation. We also established an efficient method of rAd infection to mouse ES cell lines after removing feeder cells, showing that this system is useful for removal of FRT-flanked drug-resistance gene cassette from recombinant ES cells prior to introduction of ES cells into blastocytes for chimeric mice production. Because our sequential gene-regulation system offers efficient purpose-gene regulation and strict OFF-regulation, it is potentially valuable for elucidating not only novel gene functions using cDNA microarray analysis but also for "gene switching" in development and regeneration research.
引用
收藏
页码:831 / 843
页数:13
相关论文
共 47 条
[1]   Genetic dissection of a mammalian replicator in the human β-globin locus [J].
Aladjem, MI ;
Rodewald, LW ;
Kolman, JL ;
Wahl, GM .
SCIENCE, 1998, 281 (5379) :1005-1009
[2]   THE FLP RECOMBINASE OF THE 2-MU CIRCLE DNA OF YEAST - INTERACTION WITH ITS TARGET SEQUENCES [J].
ANDREWS, BJ ;
PROTEAU, GA ;
BEATTY, LG ;
SADOWSKI, PD .
CELL, 1985, 40 (04) :795-803
[3]   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
[4]   Practical range of effective dose for Cre recombinase-expressing recombinant adenovirus without cell toxicity in mammalian cells [J].
Baba, Y ;
Nakano, M ;
Yamada, Y ;
Saito, I ;
Kanegae, Y .
MICROBIOLOGY AND IMMUNOLOGY, 2005, 49 (06) :559-570
[5]   KNOCKOUT MICE - ROUND-2 [J].
BARINAGA, M .
SCIENCE, 1994, 265 (5168) :26-&
[6]   RECOMBINATION WITHIN THE YEAST PLASMID 2-MU CIRCLE IS SITE-SPECIFIC [J].
BROACH, JR ;
GUARASCIO, VR ;
JAYARAM, M .
CELL, 1982, 29 (01) :227-234
[7]   Improved properties of FLP recombinase evolved by cycling mutagenesis [J].
Buchholz, F ;
Angrand, PO ;
Stewart, AF .
NATURE BIOTECHNOLOGY, 1998, 16 (07) :657-662
[8]   Targeted gene regulation and gene ablation [J].
DeMayo, FJ ;
Tsai, SY .
TRENDS IN ENDOCRINOLOGY AND METABOLISM, 2001, 12 (08) :348-353
[9]   GENOMIC TARGETING WITH A POSITIVE-SELECTION LOX INTEGRATION VECTOR ALLOWS HIGHLY REPRODUCIBLE GENE-EXPRESSION IN MAMMALIAN-CELLS [J].
FUKUSHIGE, S ;
SAUER, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (17) :7905-7909
[10]   TIGHT CONTROL OF GENE-EXPRESSION IN MAMMALIAN-CELLS BY TETRACYCLINE-RESPONSIVE PROMOTERS [J].
GOSSEN, M ;
BUJARD, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (12) :5547-5551