Part:BBa_K4664004
Reporter
The reporter is used for showing the fluorescence signal.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Background
Our reporter is composed of a fluorophore linked to a quencher molecule. While the two are connected, the quencher absorbs energy from the fluorophore, thus preventing it from emitting fluorescence. But when cas12a trans-cleavage acts on the quencher-fluorophore and severs their link, the fluorophore would start giving out fluorescence. Thus, the fluorescence level is used to quantify the concentration of miRNA biomarkers in the sample tested.
Design
Sequence:
5-FAM-TTATT-IABkFQ-3’
Results
We performed a series of standard MB-ERC2 reactions with different reporter concentrations at miRNA concentration levels 1 pM and 10 pM. Figures 1-3 show results from these trials.
Figure 1. Reporter 1 pM.
Figure 2. Reporter 10 pM.
Figure 3. Reporter final values.
References
Jin, J., Vaud, S., Zhelkovsky, A., Pósfai, J., & McReynolds, L. A. (2016). Sensitive and specific miRNA detection method using SplintR Ligase. Nucleic Acids Research, 44(13), e116. https://doi.org/10.1093/nar/gkw399
He, Y., Wen, Y., Tian, Z., Hart, N. T., Han, S., Hughes, S. J., & Zeng, Y. (2023b). A one-pot isothermal Cas12-based assay for the sensitive detection of microRNAs. Nature Biomedical Engineering. https://doi.org/10.1038/s41551-023-01033-1
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