Difference between revisions of "Part:BBa K3806016"
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| − | + | ==Usage and Biology== | |
| + | Sensing small molecules is the foundation of many applications, ranging from disease diagnosis, prognosis, or treatment to detecting small pollutants in the environment. Synthetic genetic switches are a promising tool to detect and quantify small molecules. Conventional synthetic biology-based biosensors are based on transcription factors. However, transcription factors are not easily reprogrammable with respect to ligand selectivity. Antibody-based biosensors can be easily developed into new sensing capabilities. Unfortunately, these types of sensors are not suitable to detect low molecular weight compounds. RNA-based biosensors are an interesting alternative since they present a remarkable flexibility to be engineered into sensing a wide range of analytes with high sensitivity and selectivity. In particular, aptazymes, ligand regulated self-cleaving ribozymes, are of special interest as cleavage of the aptazyme can be coupled to regulated gene expression in vivo or in vitro. Moreover, newly developed methods such as DRIVER (de novo rapid in vitro evolution of RNA biosensors) enable rapid, automated, and multiplexed engineering of aptazymes sequences to diverse ligands. | ||
| + | The TU Delft 2021 team provides the iGEM community with a novel genetic switch construct (<a href="https://parts.igem.org/Part:BBa_K3806016" target="_blank"><b>BBa_K3806016</b></a>) in which an aptazyme sequence is fused to the <i>lacZ</i> reporter gene to convert a ligand concentration to a colorimetric read-out. The <a href="https://parts.igem.org/Part:BBa_K3806016" target="_blank"><b>BBa_K3806016</b></a> contains a theophylline-binding aptazyme sequence [1], yet this part is designed to be modular and serve as a template to engineer other ligand-specific genetic circuits by swapping the aptazyme domain. This part was successfully expressed in a cell-free system, and its biosensing performance was compared to <a href="https://parts.igem.org/Part:BBa_K3806014" target="_blank"><b>BBa_K3806014</b></a>, showing an improvement in the dynamic range and time-response capabilities. | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 22:21, 20 October 2021
Theophylline-binding aptazyme regulating lacZ expression (semi-cRBS). With T7 promoter.
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Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3204
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2321
- 1000COMPATIBLE WITH RFC[1000]