Part:BBa_K4245201

hsa-miR-1-3p RCT Padlock Probe This part is the sequence for the hsa-miR-1-3p RCT Padlock Probe, which our team designed to be specific to BBa_K4245006, hsa-miR-1-3p. This miRNA acts as a biomarker for Coronary Artery Disease, and is therefore potentially useful for the early detection of this condition. When using the Rolling Circle Transcription approach, the produced transcript contains repeats of the middle sequence (hsa-miR-1-3p, the broccoli fluorescent RNA aptamer, and the padlock spacer sequence)

Figure 1. The rolling circle transcription approach applied to our padlock probe.

When the target miRNA is present, the miRNA and the padlock probe arms form a DNA-RNA hybridization. The introduction of SplintR ligase circularizes the padlock probe, allowing T7 RNA polymerase to attach and begin transcription (Wang et al., 2014). The product is a long, single-stranded RNA transcript that contains many copies of the reverse complement of the template strand, the padlock probe (Clausson et al., 2015). (see Fig. 1).
References Clausson, C.-M., Arngården, L., Ishaq, O., Klaesson, A., Kühnemund, M., Grannas, K., Koos, B., Qian, X., Ranefall, P., Krzywkowski, T., Brismar, H., Nilsson, M., Wählby, C., & Söderberg, O. (2015). Compaction of rolling circle amplification products increases signal integrity and signal-to-noise ratio. Scientific Reports, 5(1). https://doi.org/10.1038/srep12317 Wang, X., Li, C., Gao, X., Wang, J., & Liang, X. (2015). Preparation of small RNAS using rolling circle transcription and site-specific RNA disconnection. Molecular Therapy - Nucleic Acids, 4. https://doi.org/10.1038/mtna.2014.66