Part:BBa_K4432302
PANTR1 Toehold Switch Trigger 02 Transcriptional Unit
This part is a transcription unit of the PANTR1 Trigger N°2 (BBa_K4432202) for sequence-based detection of PANTR1, a long non coding (lnc) RNA overexpressed in different types of cancer cells.
The transcription is controlled by the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000).
Usage and Biology
A toehold switch [1] (Figure 1) is an RNA-based device is composed of several parts which are essential for the efficient functioning of our biosensor :
- a Trigger Binding Site (TBS) which is a sequence complementary to the RNA target to be detected
- a Ribosome Binding Site (RBS) which allows the start of the translation only when the toehold switch is unfolded
- a start codon AUG which constitutes the start of the translation
- a linker which constitutes a bridge between the Start codon and our gene of interest
- a repressed gene of interest that is translated into a protein only when the hairpin is unfolded and the RBS exposed
The functioning comes as follows :
The binding between the RNA biomarker and the toehold switch unfolds the hairpin structure. The RBS becomes accessible now so that the translation can take place, and the initially repressed gene of interest can then be translated into a protein.
Figure 1. Principle of the toehold switch (inspired from iGEM Evry Paris-Saclay 2020).
This part is a transcription unit of BBa_K4432202 which is a trigger for the PANTR1 Toehold Switch N°2 (BBa_K4432002).
This part proved not to be functional: the transcribed PANTR1 lncRNA fragment was not able to bind to the corresponding toehold switches and no readable output was generated.
Full results are available on the BBa_K4432102 page in the registry.
References
[1] Green AA, Silver PA, Collins JJ, Yin P. Toehold switches: de-novo-designed regulators of gene expression. Cell (2014) 159, 925-939.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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