Drug-inducible RNA aptaswitch for regulating eukaryotic protein expression
Using in vivo evolution techniques, scientists in the Liu laboratory have engineered a drug-inducible aptaswitch for regulating eukaryotic protein expression at the transcription level. This aptaswitch consists of two modular RNA elements: a transcriptional activating RNA (an evolved RNA based transcriptional activators with potency comparable to that of the most active naturally occurring protein transcriptional activation domains; see reference #1) and a tetramethylrosamine (TMR)-binding aptamer. Binding of TMR, a cell-permeable small molecule, induces a conformation change, and the RNA transcriptional activation domain displays 10-fold higher activity which increases transcription of the target gene. The method of generating aptamers to a protein target of interest and appending a small molecule binding aptamer may serve as a general approach to creating small molecule-dependent regulators of biological function in living cells.
Applications
Biological small-molecule-dependent switches sense external chemical signals and transduce them into appropriate internal signals and cellular responses. Artificial molecular switches that control the function of any protein of interest using a small molecule are powerful tools for studying biology because they enable cellular responses to be controlled by inputs chosen by researcher. Furthermore, these switches can combine the generality of genetic regulation with the reversibility and temporal control afforded by small molecules.
Using in vivo evolution techniques, scientists in the Liu laboratory have engineered a drug-inducible aptaswitch for regulating eukaryotic protein expression at the transcription level. This aptaswitch consists of two modular RNA elements: a transcriptional activating RNA (an evolved RNA based transcriptional activators with potency comparable to that of the most active naturally occurring protein transcriptional activation domains; see reference #1) and a tetramethylrosamine (TMR)-binding aptamer. Binding of TMR, a cell-permeable small molecule, induces a conformation change, and the RNA transcriptional activation domain displays 10-fold higher activity which increases transcription of the target gene. The method of generating aptamers to a protein target of interest and appending a small molecule binding aptamer may serve as a general approach to creating small molecule-dependent regulators of biological function in living cells.
Biological small-molecule-dependent switches sense external chemical signals and transduce them into appropriate internal signals and cellular responses. Artificial molecular switches that control the function of any protein of interest using a small molecule are powerful tools for studying biology because they enable cellular responses to be controlled by inputs chosen by researcher. Furthermore, these switches can combine the generality of genetic regulation with the reversibility and temporal control afforded by small molecules.
Intellectual Property Status: Patent(s) Pending