Difference between revisions of "Part:BBa K3431022"

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<partinfo>BBa_K3431022 short</partinfo>
 
<partinfo>BBa_K3431022 short</partinfo>
  
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===Description===
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===Introduction===
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Toehold Switch with Invertase as Expression Protein (oz21_A) 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 (oz21_A) (BBa_K3431006) - 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 ensure that our device can transcribe and translate in the environment of PURExpress in vitro protein synthesis kit.
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oz21_A_ToeholdSwitch-Regulated Invertase is a genetic device that can be
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applied as a biosensor for miRNA. It is designed to detect and measure the
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amount of miR-21 by the expression of Thermotoga maritima Invertase
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(BBa_K3431000). The invertase can convert sucrose to glucose, which can
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be easily measured by a personal glucose meter (PGM).
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===Components===
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oz21_A_ToeholdSwitch-Regulated Invertase consists of 4 basic parts: T7 promoter (BBa_I719005), oz21_A toehold switch (BBa_K3431006), invertase
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(BBa_K3431000), and T7 terminator (BBa_K731721). The mechanism of the detection is mainly based on the regulatory part, oz21_A Toehold Switch for miR-21 Detection (BBa_K3431006). 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 process of the downstream reporter, invertase (BBa_K3431000). As for the T7 promoter (BBa_I719005) and T7 terminator (BBa_K731721), they are the essential genetic elements for the PURExpress protein synthesis kit.
  
 
===Experiment result===
 
===Experiment result===

Revision as of 13:26, 25 October 2020


oz21_A_ToeholdSwitch-Regulated Invertase

Introduction

oz21_A_ToeholdSwitch-Regulated Invertase is a genetic device that can be applied as a biosensor for miRNA. It is designed to detect and measure the amount of miR-21 by the expression of Thermotoga maritima Invertase (BBa_K3431000). The invertase can convert sucrose to glucose, which can be easily measured by a personal glucose meter (PGM).

Components

oz21_A_ToeholdSwitch-Regulated Invertase consists of 4 basic parts: T7 promoter (BBa_I719005), oz21_A toehold switch (BBa_K3431006), invertase (BBa_K3431000), and T7 terminator (BBa_K731721). The mechanism of the detection is mainly based on the regulatory part, oz21_A Toehold Switch for miR-21 Detection (BBa_K3431006). 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 process of the downstream reporter, invertase (BBa_K3431000). As for the T7 promoter (BBa_I719005) and T7 terminator (BBa_K731721), they are the essential genetic elements for the PURExpress 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
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1434
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1205
    Illegal BamHI site found at 1335
    Illegal XhoI site found at 1406
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1006
  • 1000
    COMPATIBLE WITH RFC[1000]