RNA
miR-451a

Part:BBa_K4245012

Designed by: Akshaya Poonepalle, Michelle Jing, Shivaek Venkateswaran, Daeun Lee, Sahana Ram Narayanan, Richard Jiang, Sishnukeshav Balamurali, Christina Yi, Varnica Basavaraj, Manaswi Gorle, Janet Standeven   Group: iGEM22_Lambert_GA   (2022-09-22)

hsa-miR-451a

This part is the sequence for hsa-miR-451a, an miRNA isolated from Homo sapiens. This miRNA is not linked to coronary artery disease (C. Searles, personal communication, August 24, 2022). Dr. Searles informed us that miR-451a is mostly found in erythrocytes and is one of the most abundant extracellular miRNAs in plasma or serum. As such, miR-451a levels should be consistent between samples (Mussbacher et al., 2020). Therefore, Lambert iGEM used hsa-miR-451a as a control since it is not upregulated or downregulated in the presence of coronary artery disease, and it is found in higher and consistent amounts in patients. Thus, this biomarker acts as a baseline of comparison for the other two miRNAs we gathered data on this year, hsa-miR-1-3p (BBa_K4245006) hsa-miR-133a-3p (BBa_K4245009).

The Lambert_GA 2022 team developed a padlock probe to be used with the rolling circle amplification approaches for several miRNA-451a in order to act as our control, as aforementioned. This miRNA is used as the basis for BBa_K4245106 and BBa_K4245113, the 3' arm for hsa-miR-451 and 5' arm for hsa-miR-451a, and as the target sequence for BBa_K4245209 the hsa-miR-451a RCA Padlock Probe.

When using rolling circle amplification (RCA), the miRNA binds to the padlock. A rolling circle product (RCP) is produced from BBa_K4245131 (Middle Sequence), which is then detected by the linear probes BBa_K4245130 (Fluorophore) and BBa_K4245132 (Quencher). When these parts bind to the RCP, the fluorescence decreases. Therefore, lower fluorescence is indicative of greater miRNA concentrations.

References:

Mussbacher, M., Krammer, T. L., Heber, S., Schrottmaier, W. C., Zeibig, S., Holthoff, H.-P., Pereyra, D., Starlinger, P., Hackl, M., & Assinger, A. (2020, August 18). Impact of anticoagulation and sample processing on the quantification of human blood-derived microrna signatures. MDPI. Retrieved October 10, 2022, from https://doi.org/10.3390/cells9081915

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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