Part:BBa_K3431029
or146_A_ToeholdSwitch-Regulated Invertase
Toehold Switch with Invertase as Expression Protein (or146_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-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 (or146_A) (BBa_K3431013) - 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
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 40
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 40
Illegal NheI site found at 1434 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 40
Illegal BglII site found at 1205
Illegal BamHI site found at 1335
Illegal XhoI site found at 1406 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 40
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 40
Illegal AgeI site found at 1006 - 1000COMPATIBLE WITH RFC[1000]
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