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A designed curved DNA segment that is a remarkable activator of eukaryotic transcription
Author(s) -
Sumida Noriyuki,
Nishikawa Junichi,
Kishi Haruka,
Amano Miho,
Furuya Takayo,
Sonobe Haruyuki,
Ohyama Takashi
Publication year - 2006
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2006.05557.x
Subject(s) - microbiology and biotechnology , biology , transcription (linguistics) , transgene , promoter , transfection , gene , dna , activator (genetics) , thymidine kinase , scaffold/matrix attachment region , transcription factor , gene expression , herpes simplex virus , virus , genetics , philosophy , linguistics , chromatin remodeling
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.