Composite

Part:BBa_K4245205:Design

Designed by: Michelle Jing, Daeun Lee, Richard Jiang, Sishnukeshav Balamurali, Christina Yi, Janet Standeven   Group: iGEM22_Lambert_GA   (2022-10-09)
Revision as of 17:06, 10 October 2022 by Sriragt (Talk | contribs) (→‎Design Notes)


hsa-mir-133a-3p RCT Padlock Probe


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]


Design Notes

A padlock probe, which is often 30-150 nucleotides in length, is a single-stranded DNA sequence designed to recognize a specific target sequence. The “arms” of a padlock probe are the ends of the ssDNA that are complementary to a specific target sequence. The middle sequence (the sequence between the arms) can be specifically designed to perform a function once amplified.

Lambert iGEM found three specific miRNAs — hsa-miR-1-3p (BBa_K4245006), hsa-mir-133a-3p (BBa_K4245009), and hsa-miR-208a-3p (BBa_K4245011) — to be upregulated in correlation to CAD (Kaur et al., 2020). For miRNA 1, we designed two complementary arms, BBa_K4245100, the 3' arm for hsa-miR-1-3p and BBa_K4245107, the 5' arm for hsa-miR-1-3p. For the reporter, we decided on BBa_K3380153, the Broccoli fluorescent RNA aptamer, for its short length and function similar to GFP. In order to stabilize the folding of the Broccoli fluorescent RNA aptamer, we added BBa_K4245160, the padlock spacer sequence.

Source

hsa-mir-133a-3p RCT Padlock Probe

References