کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1993662 | 1064697 | 2012 | 7 صفحه PDF | دانلود رایگان |
Artificial RNA riboswitches – apart from protein-based gene regulation systems, which have been known about for a long time – have become increasingly important in biotechnology and synthetic biology. Aptamer-controlled hammerhead ribozymes (so-called aptazymes) have been shown to be a versatile platform for the engineering of novel gene regulators. Since aptazymes are cis-acting elements that are located in the untranslated regions of a gene of interest, their application does not need any further protein co-factor. This presents the opportunity to simplify complex gene networks while simultaneously expanding the repertoire of available parts. Nevertheless, the generation of novel aptazymes requires a functional aptamer–ribozyme connection, which can be difficult to engineer. This article describes a novel approach for using fluorescence activated cell sorting (FACS) in order to identify functional aptazymes in bacteria and their subsequent transfer into mammalian cells.
Figure optionsDownload as PowerPoint slideHighlights
► Description of a novel theophylline-responsive aptazyme.
► FACS-mediated screening for functional variants in bacteria.
► Introduction of aptazymes into mammalian cells.
Journal: Methods - Volume 56, Issue 3, March 2012, Pages 351–357