Difference between revisions of "Part:BBa K3431033"
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translation of the downstream reporter. | translation of the downstream reporter. | ||
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| + | The design of the toehold switch was mainly based on the previous research. 1 2 3 4 5 6 For the op21_B toehold switch, we adopted the loop structure from Green et al., 2014 7, and the linker structure is from Wang et al., 2019 7 . Using NUPACK analysis and Vienna binding models, we designed the sequence of the toehold switch. (See our model page: | ||
| + | https://2020.igem.org/Team:CSMU_Taiwan/Model ) | ||
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| − | + | <figure style="mirgin-right: 1em; float:left; width:40%; border:1px solid black"> | |
| − | + | <img src="https://static.igem.org/mediawiki/parts/8/8e/T--CSMU_Taiwan--op21_B_NU.png" style="display: block;margin-left: auto;margin-right: auto; width: 70%"> | |
| − | <img src="https://static.igem.org/mediawiki/parts/8/8e/T--CSMU_Taiwan--op21_B_NU.png" style="width: | + | <figcaption style="text-align: center;"> |
| + | Figure 1. NUPACK analysis result | ||
| + | </figcaption> | ||
| + | </figure> | ||
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| − | + | <img src="https://static.igem.org/mediawiki/parts/c/c7/T--CSMU_Taiwan--op21_B_Ve.png" style="display: block;margin-left: auto;margin-right: auto; width: 100%"> | |
| − | + | <figcaption style="text-align: center;"> | |
| − | + | Figure 2. ViennaRNA Package result | |
| − | + | </figcaption> | |
| − | <img src="https://static.igem.org/mediawiki/parts/c/c7/T--CSMU_Taiwan--op21_B_Ve.png " style="width: | + | </figure> |
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===Experiment result=== | ===Experiment result=== | ||
Revision as of 13:54, 25 October 2020
op21_B Toehold Switch for miR-21 Detection
Introduction
op21_B toehold switch is a regulatory part for the downstream reporter gene. With this part, the protein expression can be controlled by the miR-21. The sequence of the toehold switch can be separated into the following 5 regions from its 5' end: TBS (trigger binding site), stem region, loop region with RBS (ribosome binding site), complimentary stem region with a start codon, and linker. Upon binding with miR-21, its hairpin structure can be opened up and the ribosomes can bind with its RBS (ribosome binding site), triggering the translation of the downstream reporter.
Design
The design of the toehold switch was mainly based on the previous research. 1 2 3 4 5 6 For the op21_B toehold switch, we adopted the loop structure from Green et al., 2014 7, and the linker structure is from Wang et al., 2019 7 . Using NUPACK analysis and Vienna binding models, we designed the sequence of the toehold switch. (See our model page: https://2020.igem.org/Team:CSMU_Taiwan/Model )
Experiment result
References
Green, A. A., Silver, P. A., Collins, J. J., & Yin, P. (2014). Toehold switches: de-novo-designed regulators of gene expression. Cell, 159(4), 925-939. Pardee, K., Green, A. A., Takahashi, M. K., Braff, D., Lambert, G., Lee, J. W., ... & Daringer, N. M. (2016). Rapid, low-cost detection of Zika virus using programmable biomolecular components. Cell, 165(5), 1255-1266. Wang, S., Emery, N. J., & Liu, A. P. (2019). A novel synthetic toehold switch for microRNA detection in mammalian cells. ACS synthetic biology, 8(5), 1079-1088.
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]