Kinetics of drug selection systems in mouse embryonic stem cells

Yuhki Nakatake, Setsuko Fujii, Shinji Masui, Toshimi Sugimoto, Satomi Torikai-Nishikawa, Kenjiro Adachi, Hitoshi Niwa

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: Stable expression of transgenes is an important technique to analyze gene function. Various drug resistance genes, such as neo, pac, hph, zeo, bsd, and hisD, have been equally used as selection markers to isolate a transfectant without considering their dose-dependent characters.Results: We quantitatively measured the variation of transgene expression levels in mouse embryonic stem (mES) cells, using a series of bi-cistronic expression vectors that contain Egfp expression cassette linked to each drug resistant gene via IRES with titration of the selective drugs, and found that the transgene expression levels achieved in each system with this vector design are in order, in which pac and zeo show sharp selection of transfectants with homogenously high expression levels. We also showed the importance of the choice of the drug selection system in gene-trap or gene targeting according to this order.Conclusions: The results of the present study clearly demonstrated that an appropriate choice of the drug resistance gene(s) is critical for a proper design of the experimental strategy.

Original languageEnglish
Article number64
JournalBMC Biotechnology
Volume13
DOIs
Publication statusPublished - 2013 Aug 7

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Keywords

  • Expression
  • Gene targeting
  • Marker
  • Transgene
  • Vector

ASJC Scopus subject areas

  • Biotechnology

Cite this

Nakatake, Y., Fujii, S., Masui, S., Sugimoto, T., Torikai-Nishikawa, S., Adachi, K., & Niwa, H. (2013). Kinetics of drug selection systems in mouse embryonic stem cells. BMC Biotechnology, 13, [64]. https://doi.org/10.1186/1472-6750-13-64