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===Description===
 
Toehold Switch with Invertase as Expression Protein (zr146_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-146 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-146 (zr146_B) (BBa_K3431035) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-146. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to ensure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit.
 
Toehold Switch with Invertase as Expression Protein (zr146_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-146 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-146 (zr146_B) (BBa_K3431035) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-146. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to ensure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit.
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===Experiment result===
 
===Experiment result===
 
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Revision as of 17:02, 24 October 2020


zr146_B_ToeholdSwitch-Regulated Invertase

Description

Toehold Switch with Invertase as Expression Protein (zr146_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-146 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-146 (zr146_B) (BBa_K3431035) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-146. Furthermore, we include T7 promoter (BBa_I719005) and terminator (BBa_K731721) to ensure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit.

Experiment result



Reference

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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 40
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 40
    Illegal NheI site found at 1421
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 40
    Illegal BglII site found at 1192
    Illegal BamHI site found at 1322
    Illegal XhoI site found at 1393
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 40
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 40
    Illegal AgeI site found at 993
  • 1000
    COMPATIBLE WITH RFC[1000]