Difference between revisions of "Part:BBa K3431016"
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<partinfo>BBa_K3431016 short</partinfo> | <partinfo>BBa_K3431016 short</partinfo> | ||
+ | ===Description=== | ||
Toehold Switch with Invertase as Expression Protein (pp21_B) is an RNA-based device that is to apply as a biosensor for miRNA. This biosensor is designed to detect and reflect the amount of miRNA-21 through the expression of Beta-Fructosidase (Thermotoga Maritima MSB8) (BBa_K3431000), which can convert sucrose into glucose in a time-saving process and its result can be easily represented in the readout of glucose meter. The mechanism for detection relied on the following part - Toehold Switch for miRNA-21 (pp21_B) (BBa_K3431001) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-21. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to insure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit. | Toehold Switch with Invertase as Expression Protein (pp21_B) is an RNA-based device that is to apply as a biosensor for miRNA. This biosensor is designed to detect and reflect the amount of miRNA-21 through the expression of Beta-Fructosidase (Thermotoga Maritima MSB8) (BBa_K3431000), which can convert sucrose into glucose in a time-saving process and its result can be easily represented in the readout of glucose meter. The mechanism for detection relied on the following part - Toehold Switch for miRNA-21 (pp21_B) (BBa_K3431001) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-21. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to insure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit. | ||
+ | |||
+ | ===Experiment result=== | ||
+ | <html> | ||
+ | <br> | ||
+ | <div style="width=100%; display:flex; align-items: center; justify-content: center"> | ||
+ | <img src="https://static.igem.org/mediawiki/parts/e/e5/T--CSMU_Taiwan--pp21_B_%28BBa_K3431001%29.png" style="width:50%"> | ||
+ | </div> | ||
+ | <br> | ||
+ | </html> | ||
+ | |||
+ | ===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 | ||
+ | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here |
Revision as of 16:22, 24 October 2020
pp21_B_ToeholdSwitch-Regulated Invertase
Description
Toehold Switch with Invertase as Expression Protein (pp21_B) is an RNA-based device that is to apply as a biosensor for miRNA. This biosensor is designed to detect and reflect the amount of miRNA-21 through the expression of Beta-Fructosidase (Thermotoga Maritima MSB8) (BBa_K3431000), which can convert sucrose into glucose in a time-saving process and its result can be easily represented in the readout of glucose meter. The mechanism for detection relied on the following part - Toehold Switch for miRNA-21 (pp21_B) (BBa_K3431001) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-21. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to insure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit.
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]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1419
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1190
Illegal BamHI site found at 1320
Illegal XhoI site found at 1391 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 991
- 1000COMPATIBLE WITH RFC[1000]