title: Kinetics of drug selection systems in mouse embryonic stem cells.
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.
authors.affiliation: Laboratory for Pluripotent Stem Cell Studies, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan.
authors.forename: Yuhki
authors.initials: Y
authors.lastname: Nakatake
authors.forename: Setsuko
authors.initials: S
authors.lastname: Fujii
authors.forename: Shinji
authors.initials: S
authors.lastname: Masui
authors.forename: Toshimi
authors.initials: T
authors.lastname: Sugimoto
authors.forename: Satomi
authors.initials: S
authors.lastname: Torikai-Nishikawa
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Adachi
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Niwa
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods; D004351:Drug Resistance / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D018390:Gene Targeting / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D012641:Selection, Genetic; D019076:Transgenes
chemical_list: D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
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