Part:BBa_K3431025

or31_ToeholdSwitch-Regulated Invertase

Toehold Switch with Invertase as Expression Protein (or31) 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-31 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-31 (or31) (BBa_K3431009) - whose restriction on the expression of invertase can be liberated upon binding with miRNA-31. 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
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1433
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1204
  Illegal BamHI site found at 1334
  Illegal XhoI site found at 1405
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1005
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 536