Difference between revisions of "Part:BBa K3431016"

m
Line 3: Line 3:
 
<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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1419
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1190
    Illegal BamHI site found at 1320
    Illegal XhoI site found at 1391
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 991
  • 1000
    COMPATIBLE WITH RFC[1000]