Engineering of ribozyme-based riboswitches for mammalian cells.
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.
ETH Zurich, Department of Biosystems Science and Bioengineering (D-BSSE), Mattenstrasse 26, CH-4058 Basel, Switzerland.
Markus
M
Wieland
David
D
Ausländer
Martin
M
Fussenegger
D000595:Amino Acid Sequence; D000818:Animals; D052157:Aptamers, Nucleotide / Q000235:genetics; D005818:Genetic Engineering; D006801:Humans; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000502:physiology*; D058928:Riboswitch
D052157:Aptamers, Nucleotide; D016337:RNA, Catalytic; D058928:Riboswitch; C427204:hammerhead ribozyme