Stable Expression of Antisense Rb-1 RNA Inhibits Terminal Differentiation of Mouse Myoblast C2 Cells

Kobayashi, Masayuki; Yamauchi, Yukika; Tanaka, Akiko

doi:10.1006/excr.1997.3880

Cited by 14 publications

(15 citation statements)

References 40 publications

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“…Secondly, ectopic expression of pocket proteins, and particularly of pRB, in certain cell types induces either morphological dierentiation or transcriptional activation of tissue-speci®c genes, for instance, the above mentioned¯at cell morphology in osteosarcoma cells (Sellers et al, 1998), muscle or adipocyte dierentiation by re-introduction of pocket proteins in pRB 7/7 ®broblasts (Gu et al, 1993;Schneider et al, 1994;Chen et al, 1996a;Novitch et al, 1996), neuronal dierentiation of PC12 cells (Kranenburg et al, 1995), and keratinocyte differentiation markers in HaCaT cells (Paramio et al, 1998). Moreover, pRB anti-sense oligonucleotides block in vitro muscle dierentiation of C2 myoblasts (Kobayashi et al, 1998). Thirdly, pocket proteins, pRB being the family member most extensively analysed, have been found to associate directly with a number of proteins with tissue-speci®c activities (Table 1).…”

Section: Control Of Dierentiation Through Regulation Of Cell Type-spementioning

confidence: 99%

Role of the retinoblastoma protein family, pRB, p107 and p130 in the negative control of cell growth

1998

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The retinoblastoma family of proteins, also known as pocket proteins, includes the product of the retinoblastoma susceptibility gene and the functionally and structurally related proteins p107 and p130. Pocket proteins control growth processes in many cell types, and this has been linked to the ability of pocket proteins to interact with a multitude of cellular proteins that regulate gene expression at various levels. By regulating gene expression, pocket proteins control cell cycle progression, cell cycle entry and exit, cell dierentiation and apoptosis. This review will focus on the mechanisms of regulation of pocket proteins and how modulation of pocket protein levels and phosphorylation status regulate association with their cellular targets. The coordinated regulation of pocket proteins provides the cells with a competence mechanism for passage through certain cell growth and dierentiation transitions.

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Section: Control Of Dierentiation Through Regulation Of Cell Type-spementioning

confidence: 99%

Role of the retinoblastoma protein family, pRB, p107 and p130 in the negative control of cell growth

1998

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“…The cells were routinely maintained with Dulbecco's modified Eagle's medium (4.5 g/L glucose, Sigma, St. Louis, MO, USA) supplemented with 2 mmol/L glutamine and 10% fetal calf serum (FCS, Biowest, Nuaille, France). NT2/ D1 cells were transfected with the FLAG-tagged TPRX1-expressing vector by the lipofection method, and stable transfectants were obtained by cultivation with puromycin (0.25 mg/mL, Sigma), as reported previously [13,19]. Differentiation of NT2/D1 cells was induced by cultivation with medium containing retinoic acid (10 ¡6 mol/L, all-trans type, Sigma).…”

Section: Construction Of the Tprx1 Expression Vectormentioning

confidence: 99%

Exogenous TPRX1 homeoprotein modulates the gene expression of lineage-specific transcription factors in human embryonal carcinoma cells

Mori

Sakuraoka

Suzuki

et al. 2018

Biotechnology & Biotechnological Equipment

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We reported previously that EGAM1 homeoproteins (EGAM1 and EGAM1N), transcribed from the Crxos gene as splicing variants, are expressed in preimplantation mouse embryos and mouse embryonic stem (ES) cells. Exogenous expression of these proteins affects the maintenance of an undifferentiated state and the progression of differentiation in mouse ES cells. Human tetrapeptide-repeat homeobox 1 (TPRX1), a member of the eutherian totipotent cell homeobox (ETCHbox) genes, is an ortholog of Crxos. However, the roles of TPRX1 in the differentiation of human pluripotent cells are still unknown. Because the TPRX1 transcripts were undetectable in an undifferentiated state and during the progression of differentiation in wild-type human embryonal carcinoma NT2/D1 cells, it would be advantageous to clarify the relationship between the exogenous expression of TPRX1 and the induction of genes encoding lineage-specific transcription factors in pluripotent cells. The expression of GATA6 and FOXA2, crucial transcription factors for the formation of the primitive endoderm, was upregulated, whereas that of CDX2, a crucial transcription factor for the formation of the trophectoderm, was downregulated by enforced expression of TPRX1. Overall, we suggest that TPRX1 is capable of modulating the expression of lineage-specific transcription factors in pluripotent cells derived from humans.

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“…Antisense Rb RNA expression vector pMK1059/ AS3'UTR14x2, directed toward a portion of the 3'UTR of Rb-1 mRNA (3'UTR14x2), was constructed by use of pMK10 59 as described [8,10].…”

Section: Construction Of Expression Vectorsmentioning

confidence: 99%

“…Earlier we constructed an effective antisense Rb RNA expression vector, pMK1059/AS3'UTR14x2 [8]. This vector expresses a single mRNA concomitantly encoding duplicated antisense Rb sequences directed to a portion of the 3'-untranslated region (3'UTR) of Rb-1 mRNA [9], an internal ribosomal e n t r y s i t e ( I R E S ) d e r i v e d f r o m t h e encephalomyocarditis virus, and the neomycinresistance gene (Neo).…”

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confidence: 99%

“…In cells transfected with pMK1059/AS3'UTR14x2, the secondary structure of the IRES in the transcript is recognized by ribosomes, and the Neo placed downstream of the IRES is translated IRES-dependently [10]. The structure of the antisense vector is advantageous to ensure expression of the antisense RNA in the majority of cells expressing a selectable marker gene [8,11].…”

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confidence: 99%

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Antisense p53 RNA Abrogates Antisense Rb RNA-Induced Cell Death in Mouse Fibroblast SV-T2 Cells.

小林

山内

田仲³

1999

Journal of Reproduction and Development

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Abstract. To directly examine the cellular roles of the retinoblastoma gene product (pRb), we constructed an effective antisense Rb RNA vector that expressed an antisense Rb-1 sequence directed toward the 3' untranslated region of Rb-1 mRNA. In this study, we introduced the antisense Rb vector into mouse fibroblast SV-T2 cells expressing large amounts of SV40 LT, which inhibits the biological functions of pRb through direct binding. Only a small number of drugresistant colonies were obtained after drug selection. Co-transfection of SV-T2 cells with pRb expression vectors and the antisense Rb vector resulted in an apparent increase in the number of drug-resistant colonies, suggesting that a decrease in cellular pRb content induced cell death in the cells. The cell death in SV-T2 cells was also abrogated by simultaneous transfection with antisense p53 RNA expression vector. These results indicate that a decrease in the amount of biologically active pRb by both the effects of the transfected antisense Rb RNA and the large amount of endogenous SV40 LT induces cell death in SV-T2 cells through a p53-dependent pathway. The antisense Rb and p53 vectors used in this study are useful to examine the biological functions of these genes in established cell lines.

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Stable Expression of Antisense Rb-1 RNA Inhibits Terminal Differentiation of Mouse Myoblast C2 Cells

Cited by 14 publications

References 40 publications

Role of the retinoblastoma protein family, pRB, p107 and p130 in the negative control of cell growth

Role of the retinoblastoma protein family, pRB, p107 and p130 in the negative control of cell growth

Exogenous TPRX1 homeoprotein modulates the gene expression of lineage-specific transcription factors in human embryonal carcinoma cells

Antisense p53 RNA Abrogates Antisense Rb RNA-Induced Cell Death in Mouse Fibroblast SV-T2 Cells.

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