Next generation biotechnology: how sophisticated constructs lead to further insights and new approaches towards biotechnology’s demands

Abstract

For most plants transformation methods and regeneration protocols are readily available, leading to more complicated and differential research in plant science. The application of expression or RNAi constructs in biotechnology can lead to unwanted effects and detrimental phenotypes, which may cause great problems in the regeneration and breeding of transgenic plants. To circumvent this set back we resort to the transformation of inactive constructs. After the creation and establishment of transgenic lines the construct is specifically activated and the effects on the plants can be studied in detail, even if the re­sulting phenotype has harmful consequences. The activation of the constructs is carried out by means of induced site-directed recombination. The application of an heat shock leads to the expression of a recombinase, which excises the spacer fragment and leads to the desired effect on the plants. The feasibi­lity of those constructs could be shown for RNAi, induced expression and even marker excision and confinement approaches demonstrating the huge variabi­lity of the new construct design.

Keywords

• Site-specific Recombination
• Cre/loxP
• FLP/FRT
• Heat Shock Inducible Promoter
• Tissue Specific Expression
• RNAi