A designed curved DNA segment that is a remarkable activator of eukaryotic transcription

Noriyuki Sumida, Jun Ichi Nishikawa, Haruka Kishi, Miho Amano, Takayo Furuya, Haruyuki Sonobe, Takashi Ohyama

    Research output: Contribution to journalArticle

    20 Citations (Scopus)

    Abstract

    To identify artificial DNA segments that can stably express transgenes in the genome of host cells, we built a series of curved DNA segments that mimic a left-handed superhelical structure. Curved DNA segments of 288 bp (T32) and 180 bp (T20) were able to activate transcription from the herpes simplex virus thymidine kinase (tk) promoter by approximately 150-fold and 70-fold, respectively, compared to a control in a transient transfection assay in COS-7 cells. The T20 segment was also able to activate transcription from the human adenovirus type 2 E1A promoter with an 18-fold increase in the same assay system, and also activated transcription from the tk promoter on episomes in COS-7 cells. We also established five HeLa cell lines with genomes containing T20 upstream of the transgene promoter and control cell lines with T20 deleted from the transgene locus. Interestingly, T20 was found to activate transcription in all the stable transformants, irrespective of the locus. This suggests that the T20 segment may allow stable expression of transgenes, which is of importance in many fields, and may also be useful for the construction of nonviral vectors for gene therapy.

    Original languageEnglish
    Pages (from-to)5691-5702
    Number of pages12
    JournalFEBS Journal
    Volume273
    Issue number24
    DOIs
    Publication statusPublished - 2006 Dec

    Fingerprint

    Transcription
    Transgenes
    Thymidine Kinase
    COS Cells
    DNA
    Assays
    Genes
    Cells
    Genome
    Human Adenoviruses
    Cell Line
    Gene therapy
    Simplexvirus
    Viruses
    HeLa Cells
    Genetic Therapy
    Transfection
    Plasmids

    Keywords

    • Chromatin
    • Chromatin engineering
    • Curved DNA
    • Supercoil
    • Transcription activator

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    A designed curved DNA segment that is a remarkable activator of eukaryotic transcription. / Sumida, Noriyuki; Nishikawa, Jun Ichi; Kishi, Haruka; Amano, Miho; Furuya, Takayo; Sonobe, Haruyuki; Ohyama, Takashi.

    In: FEBS Journal, Vol. 273, No. 24, 12.2006, p. 5691-5702.

    Research output: Contribution to journalArticle

    Sumida, N, Nishikawa, JI, Kishi, H, Amano, M, Furuya, T, Sonobe, H & Ohyama, T 2006, 'A designed curved DNA segment that is a remarkable activator of eukaryotic transcription', FEBS Journal, vol. 273, no. 24, pp. 5691-5702. https://doi.org/10.1111/j.1742-4658.2006.05557.x
    Sumida N, Nishikawa JI, Kishi H, Amano M, Furuya T, Sonobe H et al. A designed curved DNA segment that is a remarkable activator of eukaryotic transcription. FEBS Journal. 2006 Dec;273(24):5691-5702. https://doi.org/10.1111/j.1742-4658.2006.05557.x
    Sumida, Noriyuki ; Nishikawa, Jun Ichi ; Kishi, Haruka ; Amano, Miho ; Furuya, Takayo ; Sonobe, Haruyuki ; Ohyama, Takashi. / A designed curved DNA segment that is a remarkable activator of eukaryotic transcription. In: FEBS Journal. 2006 ; Vol. 273, No. 24. pp. 5691-5702.
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