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Modified S/MAR episomal vectors for stably expressing fluorescent protein-tagged transgenes with small cell-to-cell
Akiko Mizutani1, Eri Kikkawa, Akira Matsuno
1Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan; Department of Neurosurgery, Teikyo University Chiba Medical Center, Ichihara, Chiba 299-0111, Japan; Faculty of Health and Medical Science, Teikyo Heisei University, Higashi-Ikebukuro, Tokyo 170-8445, Japan.
Abstract:
We modified and tested scaffold/matrix attachment region (S/MAR) episomal vectors. The new vectors would be useful in obtaining cells stably expressing fluorescent protein-tagged transgenes with small, mostly within 10-fold cell-to-cell fluctuations. In the vectors, the same transcript directs episomal replication and expression of transgene/antibiotic marker, and only antibiotic selection without any other extra steps was sufficient to obtain desired stable cells, including those expressing two different proteins simultaneously. Furthermore, the two test cases (expression of human growth hormone in AtT20 and four protein kinase C isoforms in HEK293) would prove to be useful in visualizing and analyzing regulatory processes involving these proteins.
