Toward Higher Order Control Modalities in Mammalian Cells-Independent Adjustment of Two Different Gene Activities
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- 作者：Cornelia Fux and Martin Fussenegger
- 刊名：Biotechnology Progress
- 出版年：2003
- 出版时间：February 2003
- 年：2003
- 卷：19
- 期：1
- 页码：109 - 120
- 全文大小：535K
- 年卷期：v.19,no.1(February 2003)
- ISSN：1520-6033
文摘
Heterologous higher order control modalities will be important tools for targetedmultigene interventions in next-generation gene therapy, tissue engineering, andsophisticated gene-function studies. In this study, we present the design and rigorousquantitative analysis of a variety of different dual-regulated gene transcription controlconfigurations combining streptogramin- and tetracycline-responsive expressionsystems in a one-vector format. Quantitative assessment of dual-regulated expressionperformance in various mammalian and human cell lines is based on two compatiblesecreted reporter genes, SEAP, the human placental secreted alkaline phosphatase,and the recently developed SAMY, the secreted 
-amylase. Assembly of streptogramin-and tetracycline-responsive transgene control units in consecutive (
), divergent(
), and convergent ( ) orientation showed excellent regulation characteristicsin most genetic arrangements exemplified by neglectable interference and hightransgene induction ratios in all four control settings (ON/ON, OFF/ON, ON/OFF,OFF/OFF). The overall regulation performance of divergent dual-regulated expressionconfigurations could be substantially increased when placing noncoding stufferfragments or insulator modules between the divergently oriented antibiotic-responsivepromoters. Dual-regulated expression technology pioneers artificial higher order genecontrol networks that will likely enable new opportunities in multigene metabolicengineering and generate significant therapeutic impact.
-amylase. Assembly of streptogramin-and tetracycline-responsive transgene control units in consecutive ( ), divergent( ), and convergent ( ) orientation showed excellent regulation characteristicsin most genetic arrangements exemplified by neglectable interference and hightransgene induction ratios in all four control settings (ON/ON, OFF/ON, ON/OFF,OFF/OFF). The overall regulation performance of divergent dual-regulated expressionconfigurations could be substantially increased when placing noncoding stufferfragments or insulator modules between the divergently oriented antibiotic-responsivepromoters. Dual-regulated expression technology pioneers artificial higher order genecontrol networks that will likely enable new opportunities in multigene metabolicengineering and generate significant therapeutic impact.