Efficient gene activation system on mammalian cell chromosomes using recombinant adenovirus producing Cre recombinase

Kanegae, Yumi; Takamori, Koichi; Sato, Yuichi; Lee, Gwang; Nakai, Michiko; Saito, Izumu

doi:10.1016/s0378-1119(96)00516-1

Cited by 103 publications

(95 citation statements)

References 11 publications

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“…However, applications of the Cre-loxP system were still very limited because only a small fraction of cells could be subjected to gene activation by Cre recombinase produced as a result of DNA transfection or electroporation. Kanegae et al 28 reported that Cre-mediated gene activation could be achieved in virtually 100% of a cultured cell population on cell chromosomes by using adenoviral vectors expressing a modified Cre gene tagged with a nuclear localization signal. 28,29 Sato et al 20 established a strategy for introducing exogenous Bcl-2 into neuronal cells in vitro that is mediated by Cre-loxP recombination using adenoviral vectors.…”

Section: Figure 7 Detection Of Immunoreactivity For Bcl-2 In the Tongmentioning

confidence: 99%

Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

et al. 2001

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We investigated genes expression by retrograde axonal transport of replication-defective adenoviruses carrying genes for LacZ (AdLacZ) and Bcl-2 in motor neurons of transgenic mice expressing mutant human Cu/Zn superoxide dismutase (SOD1) gene containing a substitution of alanine for glycine at position 93. We found that intramuscular injection of AdLacZ into the tongue of mutant SOD1 transgenic mice and their wild-type littermates at various ages results in high expression of the transgene and similar time course of expression in hypoglossal cranial nerve nuclei, suggesting no difference in the behavior of the transgene expression between the two groups. Subsequently, we employed a molecular switching cassette for Bcl-2 designed to express Bcl-2 by Cre-loxP recombination using adenoviral vectors, and examined the COS7 and primary neuronal cells with the mutant SOD1 gene. The overexpression of Bcl-2 in

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Section: Figure 7 Detection Of Immunoreactivity For Bcl-2 In the Tongmentioning

confidence: 99%

Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

et al. 2001

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“…These ®ndings prompted us to examine the involvement of h-l(3)mbt in cell cycle regulation. To investigate the role of h-l(3)mbt in proliferating cells, we employed the inducible overexpression of h-l(3)mbt gene in U251MG cells by using the Cre-mediated gene activation system (Kanegae et al, 1995(Kanegae et al, , 1996. The U251MG cells, in which endogenous h-l(3)mbt proteins has not been detected, were transfected with pCALNL5, pCALNL5/h-l(3)mbt-I, or pCALNL5/ h-l(3)mbt-II, and clones were selected with G418.…”

Section: Overexpression Of H-l(3)mbt Induces Multinucleated Cellsmentioning

confidence: 99%

“…The constructs corresponding to Cre-mediated h-l(3)mbt-I and h-l(3)mbt-II expression plasmids were generated by ligating the EcoRI/BamHI fragments containing the fulllength ORF of the h-l(3)mbt-I and h-l(3)mbt-II cDNA into the pCALNL5 plasmid which has a Cre-mediated activation unit (Kanegae et al, 1995(Kanegae et al, , 1996. These constructs were named pCALNL5/h-l(3)mbt-I and pCALNL5/h-l(3)mbt-II.…”

Section: Inducible Expression Of the H-l(3)mbt In Mammalian Cellsmentioning

confidence: 99%

“…Clones were selected in medium containing 400 mg/ml geneticin (GIBCO BRL). The drug-resistant cell lines were infected with AxCANCre adenovirus which produces Cre recombinase (Kanegae et al, 1996) for 12 h in DMEM/F12 culture medium, and then the medium was changed to adenovirus-free growing medium. More than 48 h after the infection, cells were prepared for Western blot and CLSM analyses as described above.…”

Section: Inducible Expression Of the H-l(3)mbt In Mammalian Cellsmentioning

confidence: 99%

“…These ®ndings suggest that the h-l(3)mbt is structurally similar to a member of PcG chromatin-binding complex but is not involved in the previously identi®ed PcG complex. Furthermore, we expressed the h-l(3)mbt gene in the human glioma cell line U251MG with the use of an adenovirus-mediated gene activation system (Kanegae et al, 1995(Kanegae et al, , 1996, and we found that overexpression of h-l(3)mbt a ects nuclear segregation and cytokinesis, resulting in the formation of multinucleated cells. The h-l(3)mbt is potentially involved in regulation of cell cycle progression by participating in the formation of a DNA-binding protein complex.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A human homolog of Drosophila lethal(3)malignant brain tumor (l(3)mbt) protein associates with condensed mitotic chromosomes

et al. 1999

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A novel human nucleoside diphosphate (NDP) kinase, Nm23-H6, localizes in mitochondria and affects cytokinesis

et al. 2000

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Nucleoside diphosphate kinases (NDP kinases) are enzymes known to be conserved throughout evolution and have been shown to be involved in various biological events, in addition to the "housekeeping" phosphotransferase activity. We present the molecular cloning of a novel human NDP kinase gene, termed Nm23-H6. Nm23-H6 gene has been mapped at chromosome 3p21.3 and is highly expressed in heart, placenta, skeletal muscle, and some of the cancer cell lines. Recombinant Nm23-H6 protein has been identified to exhibit functional NDP kinase activity. Immunolocalization studies showed that both endogenous and inducibly expressed Nm23-H6 proteins were present as short, filament-like, perinuclear radical arrays and that they colocalized with mitochondria. Cell fractionation study also demonstrated the presence of Nm23-H6 protein in a mitochondria-rich fraction. Moreover, induction of overexpression of Nm23-H6 in SAOS2 cells, using the Cre-loxP gene activation system, resulted in growth suppression and generation of multinucleated cells. Flow cytometric analysis also demonstrated that the proportion of cells with more than 4N DNA content increased to 28.1% after induction of Nm23-H6, coinciding with the appearance of multinucleated cells. These observations suggest that Nm23-H6, a new member of the NDP kinase family, resides in mitochondria and plays a role in regulation of cell growth and cell cycle progression.

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Efficient gene activation system on mammalian cell chromosomes using recombinant adenovirus producing Cre recombinase

Cited by 103 publications

References 11 publications

Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

A human homolog of Drosophila lethal(3)malignant brain tumor (l(3)mbt) protein associates with condensed mitotic chromosomes

A novel human nucleoside diphosphate (NDP) kinase, Nm23-H6, localizes in mitochondria and affects cytokinesis

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