This RNA-based methodology circumvents the immune responses observed in other systems and achieves a 900-fold induction with an EC 50 of 0
In this setup, the presence of tetracycline inhibits miRNA-mediated transgene knockdown, thereby serving as an on-switch for gene expression
5 and 10 µM aptamer RNA was used with the tetracycline concentrations varying from 11-64 µM to 15-155 µM, respectively
1016/s0968-0896 (01)00063-3
The tetracycline aptamer was inserted into the 5' UTR of a ß-lactamase reporter gene, enabling it to control the accessibility of the ribosome binding site (RBS, Figure 2C)
In this way, when the aptamer- µM tetracycline; and DIVE_6 with 1 mM theophylline
Efficient regulation is obtained when the aptamer is inserted close to the 5′splice site (SS) with the consensus sequence of the SS located within the aptamer stem
Furthermore, the lead cleavage pattern of this RNA changed significantly upon addition of tetracycline, suggesting that cb28 changes conformation upon tetracycline binding
Among the regulatory elements that respond to external stimuli, riboswitches are a prominent and elegant example
A tetracycline aptamer was grafted onto the hammerhead ribozyme in such a way that ligand binding to the aptamers destroys a loop-loop interaction within the ribozyme The data on protein as well as on RNA level show that the tetracycline aptamer itself can be employed as an alternative exon for efficient splicing-based RNA switches
Effector-responsive ribozymes can be engineered by inserting ligand-binding RNA aptamer sequences which control the activity of the RNA catalyst 5,6,7,8
Abstract
We inserted preselected tetracycline (tc) binding aptamers into the 5'-UTR of a GFP encoding mRNA
In this study, we designed a versatile and highly efficient TetR Splicing Device (TSD) for the conditional control of gene expression in human cells that makes use of an RNA aptamer recognized by the bacterial Tet Repressor TetR
The advantage of the presented regulatory system is its independence of any regulatory proteins
This technique has been successfully employed to study essential genes in P
To study the mode of interaction of tetracycline with RNA, we isolated aptamers with high affinity to this antibiotic via in vitro selection
Tetracycline induces exon skipping resulting in ∼5-fold activation of gene expression
The 2
Small aptamer-based regulatory devices can be designed to control a range of RNA-dependent cellular processes and emerged as promising tools for fine-tuning gene expression in synthetic biology Aptamers or ribozymes could be grafted to microRNAs to control their processing and subsequent use in gene control
The design allows for an easy, context-independent Allosteric ribozymes control gene expression by cleaving RNA in response to of a theophylline aptamer with a tetracycline aptamer, showing that, at least for small molecules, the design is 10
Notably, we demonstrate diverse regulatory roles of Tamra_Ahx_R8_L-Apt
A tetracycline aptamer was grafted onto the hammerhead ribozyme in such a way that ligand binding to the aptamers destroys a loop-loop interaction within the ribozyme thereby inhibiting ribozyme cleavage and allowing In RNA-aptamer applications, RNA aptamers are used to control diverse biological functions, as well as for the construction of cellular logic gates [190][191] [192] [193]
One of the best studied laboratory-selected aptamers binds the antibiotic tetracycline, but it binds with a much lower affinity to the closely related but more bioavailable antibiotic Yen, L
Chem 9, 2549-2556 (2001)
This technique has been successfully employed to study essential genes in P
5 µg ml −1
Synthetic RNA (RNA)-based gene switches control RNA functions in a ligand-responsive manner
We apply the synthetic tetracycline (tc)-binding riboswitch to establish a gene expression system for conditional tc-dependent control of pre-mRNA splicing in
The tetracycline-binding aptamer (TC aptamer), which is in the focus of our study, was selected in vitro and optimized by a rational design approach (23, 24)
Since its selection, it has been used in a variety of applications and is
22 was introduced into the host RNA to create a tetracycline artificial riboswitch, which could control the expression of the downstream reporter gene Abstract
We developed tetracycline-dependent ribozymes, which can switch on gene expression up to 8
Researchers developed an RNA-based switch, the pA regulator system, to control gene expression in mammalian cells by modulating synthetic polyA signal cleavage, offering a novel approach for gene Two previous studies have also made use of RNAi in combination with aptamers in order to implement ligand-dependent control of gene expression in mammalian cells
We developed tetracycline-dependent ribozymes, which can switch on gene expression up to 8
Hartig, University of The tetracycline aptamer was inserted into the 5’ UTR of a ß-lactamase reporter gene, enabling it to control the accessibility of the ribosome binding site (RBS, Figure 2C)
2 Å-resolution cocrystal structure of the aptamer reveals a pseudoknot-like
Binding of the ligand often leads to a conformational change that causes structural changes of the RNA tertiary fold
We apply the synthetic tetracycline (tc)-binding riboswitch to establish a gene expression system for conditional tc-dependent control of pre-mRNA splicing in yeast
We present a conditional gene expression system in Saccharomyces cerevisiae which exploits direct RNA-metabolite interactions as a mechanism of genetic control
We present a conditional gene expression system in Saccharomyces cerevisiae which exploits direct RNA-metabolite interactions as a mechanism of genetic control
Translation in vivo was suppressed up to ninefold upon addition of tetracycline
They consist solely of RNA and couple binding of a small molecule ligand to the so-called “aptamer domain” with a conformational change in the downstream The cb28 aptamer binds tetracycline with a Kd of approximately 1 μM (see below), which is also the affinity range in which tetracycline binds to the E
We developed tetracycline-dependent ribozymes, which can switch on gene expression up to 8