Establishment of composite DNA derived from L factor as a plasmid in mouse embryonal carcinoma (F9) cells

1988 ◽  
Vol 8 (5) ◽  
pp. 2097-2104
Author(s):  
K Nishimori ◽  
T Kohda ◽  
J Fujiwara ◽  
M Oishi
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Retinoic Acid ◽  
Plasmid Dna ◽  
Inverse Relationship ◽  
Embryonal Carcinoma ◽  
Foreign Gene ◽  
In Vitro Differentiation ◽  
F9 Cells

We have recently reported a mammalian cell plasmid (L factor) whose structure is related to that of polyomavirus (T. Kusano, H. Uehara, H. Saito, K. Segawa, and M. Oishi, Proc. Natl. Acad. Sci. USA 84:1789-1793, 1987). When composite DNA constructed from L factor and a foreign gene was introduced into mouse embryonal carcinoma (F9) cells by transfection, the DNA was reestablished in the cells as a plasmid. The reestablished plasmid DNA in F9 cells could be rescued in Escherichia coli. The plasmid-bearing cells underwent normal in vitro differentiation in response to retinoic acid. The efficiency of plasmid establishment of the L-factor-derived DNA and transcriptional and transient replicational activities were compared with those of similar composite DNA constructed from polyomavirus and an embryonal carcinoma mutant of polyomavirus which is permissive in F9 cells. The results suggest an inverse relationship between the efficiency of the plasmid establishment and the activity of gene expression controlled by the intrinsic enhancer-promoter of the DNA.

scholarly journals Establishment of composite DNA derived from L factor as a plasmid in mouse embryonal carcinoma (F9) cells.

1988 ◽  
Vol 8 (5) ◽  
pp. 2097-2104 ◽  
Author(s):  
K Nishimori ◽  
T Kohda ◽  
J Fujiwara ◽  
M Oishi
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Retinoic Acid ◽  
Plasmid Dna ◽  
Inverse Relationship ◽  
Embryonal Carcinoma ◽  
Foreign Gene ◽  
In Vitro Differentiation ◽  
F9 Cells

We have recently reported a mammalian cell plasmid (L factor) whose structure is related to that of polyomavirus (T. Kusano, H. Uehara, H. Saito, K. Segawa, and M. Oishi, Proc. Natl. Acad. Sci. USA 84:1789-1793, 1987). When composite DNA constructed from L factor and a foreign gene was introduced into mouse embryonal carcinoma (F9) cells by transfection, the DNA was reestablished in the cells as a plasmid. The reestablished plasmid DNA in F9 cells could be rescued in Escherichia coli. The plasmid-bearing cells underwent normal in vitro differentiation in response to retinoic acid. The efficiency of plasmid establishment of the L-factor-derived DNA and transcriptional and transient replicational activities were compared with those of similar composite DNA constructed from polyomavirus and an embryonal carcinoma mutant of polyomavirus which is permissive in F9 cells. The results suggest an inverse relationship between the efficiency of the plasmid establishment and the activity of gene expression controlled by the intrinsic enhancer-promoter of the DNA.

Control of beta-interferon expression in murine embryonal carcinoma F9 cells

1989 ◽  
Vol 9 (8) ◽  
pp. 3553-3556
Author(s):  
M K Francis ◽  
J M Lehman
Keyword(s):  
Retinoic Acid ◽  
Embryonal Carcinoma ◽  
Biologically Active ◽  
Rnase Protection ◽  
F9 Cells ◽  
Differentiated Cells ◽  
Transcriptional Regulatory Mechanism ◽  
Transcriptional Regulatory ◽  
Tissue Culture Model

Murine embryonal carcinoma F9 cells, a tissue culture model for early embryonic development, do not produce interferon (IFN) in response to poly(I-C), as determined by an antiviral assay. RNase protection analyses were used to examine total RNA extracted from the cells for the presence of beta-IFN RNA. Whereas F9 cells differentiated in vitro with retinoic acid produced a biologically active protein as well as beta-IFN RNA in response to poly(I-C), undifferentiated F9 cells produced no detectable beta-IFN RNA even in the presence of cycloheximide, an IFN-superinducing agent. These results show that undifferentiated embryonal carcinoma cells do not accumulate beta-IFN RNA in response to an IFN-inducing agent, suggesting a transcriptional regulatory mechanism. However, this control mechanism is altered upon differentiation, since the gene can be transcriptionally activated in retinoic acid-differentiated cells.

Retinoic acid induces activin receptor IIB mRNA in F9 embryonal carcinoma cells

1995 ◽  
Vol 14 (2) ◽  
pp. 247-254 ◽  
Author(s):  
Y-J Y Wan ◽  
L Wang ◽  
T-C J Wu
Keyword(s):  
Gene Expression ◽  
Retinoic Acid ◽  
Embryonal Carcinoma ◽  
Actinomycin D ◽  
Inductive Effect ◽  
Receptor Type ◽  
Dependent Manner ◽  
Mrna Isoforms ◽  
Activin Receptor ◽  
F9 Cells

ABSTRACT Mouse embryonal carcinoma F9 cells are pluripotent stem cells and differentiate into primitive endodermal cells upon treatment with retinoic acid (RA). We have recently shown that in F9 cells RA regulates gene expression of activin receptor type II (ActR-II), whose ligand is a potent differentiation agent. The present study examined the regulation of the newly cloned activin receptor type IIB (ActR-IIB) gene by RA. F9 cells expressed equal amounts of three ActR-IIB transcripts of 8·0, 7·5 and 4·0 kb. Both 9-cis-RA (c-RA) and all-trans-RA (t-RA) induced ActR-IIB gene expression in a dose-dependent manner. At 10−9m c-RA exerted no effect, while 10−5m c-RA increased the 8·0 kb ActR-IIB transcript about sevenfold. In contrast, t-RA induced the 8·0kb ActR-IIB transcript fivefold at 10−9m and up to eightfold at 10−5m. The inductive effect on the 8·0 kb transcript was greater than that on the 7·5 kb transcript, and was least effective on the 4·0 kb transcript, suggesting that these three mRNA isoforms may originate from different promoters. Both cycloheximide and actinomycin D inhibited the inductive effect of t-RA on ActR-IIB gene expression, in contrast to ActR-II whose gene expression was not suppressed by cycloheximide but abolished by actinomycin D. Thus, endodermal differentiation of F9 cells is associated with activation of ActR-IIB gene and the mechanisms involved in the regulation of ActR-II and IIB gene expression are different.

scholarly journals Cellular HSP90 (HSP86) mRNA level and in vitro differentiation of mouse embryonal carcinoma (F9) cells

FEBS Letters ◽  
1991 ◽  
Vol 290 (1-2) ◽  
pp. 107-110 ◽  
Author(s):  
Takashi Kohda ◽  
Kazuhiro Kondo ◽  
Michio Oishi
Keyword(s):  
Embryonal Carcinoma ◽  
Mrna Level ◽  
In Vitro Differentiation ◽  
F9 Cells

scholarly journals Control of beta-interferon expression in murine embryonal carcinoma F9 cells.

1989 ◽  
Vol 9 (8) ◽  
pp. 3553-3556 ◽  
Author(s):  
M K Francis ◽  
J M Lehman
Keyword(s):  
Retinoic Acid ◽  
Embryonal Carcinoma ◽  
Biologically Active ◽  
Rnase Protection ◽  
F9 Cells ◽  
Differentiated Cells ◽  
Transcriptional Regulatory Mechanism ◽  
Transcriptional Regulatory ◽  
Tissue Culture Model

Murine embryonal carcinoma F9 cells, a tissue culture model for early embryonic development, do not produce interferon (IFN) in response to poly(I-C), as determined by an antiviral assay. RNase protection analyses were used to examine total RNA extracted from the cells for the presence of beta-IFN RNA. Whereas F9 cells differentiated in vitro with retinoic acid produced a biologically active protein as well as beta-IFN RNA in response to poly(I-C), undifferentiated F9 cells produced no detectable beta-IFN RNA even in the presence of cycloheximide, an IFN-superinducing agent. These results show that undifferentiated embryonal carcinoma cells do not accumulate beta-IFN RNA in response to an IFN-inducing agent, suggesting a transcriptional regulatory mechanism. However, this control mechanism is altered upon differentiation, since the gene can be transcriptionally activated in retinoic acid-differentiated cells.

Ultrasound/Microbubble Enhances Foreign Gene Expression in ECV304 Cells and Murine Myocardium

2004 ◽  
Vol 36 (12) ◽  
pp. 824-831 ◽  
Author(s):  
Dongping Guo ◽  
Xiaoyu Li ◽  
Ping Sun ◽  
Zhiguang Wang ◽  
Xiuying Chen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Therapy ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Ultrasound Irradiation ◽  
Ultrasound Contrast Agents ◽  
Foreign Gene ◽  
Exogenous Gene

Abstract Although viral vectors are efficient systems to transfer foreign genes into cells or target tissues, safety issues remain in relation to human gene therapy. Microbubbles currently used as ultrasound contrast agents have been applied in transfection of genes. This study was designed to test the transfection efficiency and the expression of exogenous gene mediated by ultrasound irradiation enhanced air filled albumin microbubbles in ECV304 cell line in vitro and the heart of the mouse in vivo. Air filled microbubbles (2.0–4.0 μm in diameter) were created by sonicating the mixture of human albumin, glucose, mannitol and special additive that was designed for stabilization. Plasmid DNA loading the reporter genes was gently mixed with microbubbles. The mixture of plasmid DNA and microbubbles was administrated to cultured ECV304 cells and BALB/c mice (tail vein injection) under different ultrasound/microbubble conditions, and then the transfection and expression efficiency were examined. The results both in vivo and in vitro demonstrated that microbubble with ultrasound irradiation could significantly elevate the exogenous gene expression as compared with microbubble or ultrasound only. Overall, the present study showed that the ultrasound-target microbubble destruction method enhanced the exogenous gene expression in vivo and in vitro, and provided a gene therapy way not only efficient but also easy to be manipulated and carried out in clinical.

Induction of CAT gene expression on a plasmid vector (L factor) by retinoic acid in mouse embryonal carcinoma (F9) cells

Plasmid ◽  
1991 ◽  
Vol 26 (3) ◽  
pp. 201-208 ◽  
Author(s):  
Katsuhiko Nishimori ◽  
Takashi Kohda ◽  
Kaoru Segawa ◽  
Michio Oishi
Keyword(s):  
Gene Expression ◽  
Retinoic Acid ◽  
Embryonal Carcinoma ◽  
Plasmid Vector ◽  
F9 Cells ◽  
Cat Gene
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