title: Establishment of tetracycline-inducible, survivin-expressing CHO cell lines by an optimized screening method.
abstract: An optimized method based on tetracycline-inducible gene expression system T-REx was developed to screen and evaluate Tet repressor (TetR)-expressing cell lines using enhanced green fluorescence protein (EGFP) as reporter gene. To verify the effectiveness of the method, two TetR-expressing Chinese hamster ovary (CHO) cell lines, CHO-TR B2 (stringent) and B5 (less stringent), in which the EGFP genes were variantly controlled by tetracycline, were used to construct cell lines expressing the anti-apoptosis gene survivin upon induction with tetracycline. The resulting stable clones were analyzed for survivin expression. The analysis showed that all four B5-derived clones exhibited leaky survivin expression in the absence of tetracycline, while the B2-derived clones did not. DNA laddering and annexin V/PI staining assays further indicated that although tetracycline-inducible expression of survivin conferred resistance to NH₄Cl- and staurosporine-induced apoptosis in both the B2- and the B5-derived stable cell lines, the B2-derived cell lines showed more stringent regulation in the absence of tetracycline. This represents successful utilization of the present screening method.
authors.affiliation: Beijing Institute of Biotechnology, Beijing, People's Republic of China. zhao.xinghui@gmail.com
authors.forename: Xinghui
authors.initials: X
authors.lastname: Zhao
authors.forename: Yingqun
authors.initials: Y
authors.lastname: Yu
authors.forename: Zhanzhong
authors.initials: Z
authors.lastname: Zhao
authors.forename: Junwei
authors.initials: J
authors.lastname: Guo
authors.forename: Ling
authors.initials: L
authors.lastname: Fu
authors.forename: Ting
authors.initials: T
authors.lastname: Yu
authors.forename: Lihua
authors.initials: L
authors.lastname: Hou
authors.forename: Shaoqiong
authors.initials: S
authors.lastname: Yi
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000235:genetics; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*
chemical_list: D051034:Inhibitor of Apoptosis Proteins; D012097:Repressor Proteins; C485184:enhanced green fluorescent protein; C034811:tetracycline resistance-encoding transposon repressor protein; D049452:Green Fluorescent Proteins; D013752:Tetracycline