Part:BBa_K4664008
Padlock-miR-223-3
Padlock Padlock-miR-223 is the corresponding padlock chosen for the miR-223 biomarker.
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
In order to determine the PLP sequence that yields the best reaction kinetics, we designed padlocks with the crRNA recognition site (responsible for binding with the crRNA in RNP and activating Cas12a cleavages) at different locations.
Design
In Padlock-miR-223-3（Padlock-3）, the Cas12a detection module (square brackets) is placed on the left.
Sequence:
TCAAATACACG-[CGTCGCCGTCCAGCTCGACC]-AGATACCCTAACCATCGAT-AACTCAGCTTG
Results
The following figure shows the final values for padlocks 1-4 (padlock 1 is BBa_K4664002, aka. Padlock-223).
Figure 1. Padlocks 1-4.
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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