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Competence of an artificial bent DNA as a transcriptional activator in mouse ES cells

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Abstract

Curved DNA structures with a left-handed superhelical conformation can activate eukaryotic transcription. However, their potency in transgene activation in embryonic stem (ES) cells has not been examined. T20 is an artificial curved DNA of 180 bp that serves as a transcriptional activator. We investigated the effect of T20 on transcription in mouse ES cell lines or hepatocytes differentiated from them. We established 10 sets of cell lines each harboring a single copy of the reporter construct. Each set comprised a T20-harboring cell line and a T20-less control cell line. Analyses showed that in ES cells and in hepatocytes originating from these cells, T20 both activated and repressed transcription in a manner that was dependent on the locus of reporter. The present and previous studies strongly suggest that in cells that have a strict gene regulation system, transcriptional activation by T20 occurs only in a transcriptionally active locus in the genome.

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Abbreviations

Alb :

Albumin gene

Alox5ap :

Arachidonate 5-lipoxygenase-activating protein gene

Cdk3 :

Cyclin-dependent kinase 3 gene

ChIP:

Chromatin immunoprecipitation

CK18:

Cytokeratin 18

CYC1 :

Iso-1-cytochrome c gene

Cyp7α1 :

Cholesterol 7 alpha hydroxylase gene

ES:

Embryonic stem

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GFP:

Green fluorescent protein

HSV:

Herpes simplex virus

IPCR:

Inverse polymerase chain reaction

Neo r :

Neomycin phosphotransferase gene

Oct4 :

POU domain, class 5, transcription factor 1 gene

PEPCK :

Phosphoenolpyruvate carboxykinase 1 gene

Rps18 :

Ribosomal protein S18 gene

RT-PCR:

Reverse transcription polymerase chain reaction

TAT :

Tyrosine aminotransferase gene

Tgfbr3 :

Transforming growth factor β receptor III gene

tk :

Thymidine kinase gene

UAS:

Upstream activating sequence

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Acknowledgments

The authors acknowledge the contributions of S. Fujii and Y. Kadokawa. This study was supported in part by JSPS and MEXT research grants to T.O.

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Authors and Affiliations

  1. Department of Biology, Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan

    Jun-ichi Tanase, Koji Udagawa, Jun-ichi Nishikawa & Takashi Ohyama

  2. Institute of Advanced Technology, Kinki University, 14-1 Minami-Akasaka, Kainan, Wakayama, 642-0017, Japan

    Tasuku Mitani

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  1. Jun-ichi Tanase
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  2. Tasuku Mitani
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  3. Koji Udagawa
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  4. Jun-ichi Nishikawa
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  5. Takashi Ohyama
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Corresponding author

Correspondence to Takashi Ohyama.

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Tanase, Ji., Mitani, T., Udagawa, K. et al. Competence of an artificial bent DNA as a transcriptional activator in mouse ES cells. Mol Biol Rep 38, 37–47 (2011). https://doi.org/10.1007/s11033-010-0075-5

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  • DOI: https://doi.org/10.1007/s11033-010-0075-5

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