Coding

Part:BBa_K3407007:Design

Designed by: Javier Navarro Delgado   Group: iGEM20_TUDelft   (2020-10-22)
Revision as of 10:25, 27 October 2020 by Javier (Talk | contribs)


shRNA*: short hairpin RNA with loop mutations.


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

This biobrick represents the DNA template for the transcription of a 65 nt shRNA, excluding the promoter. Trancription was performed with BBa_K3407023. The dsRNA sequence was taken from pUC57-OriLR-deGFP plasmid (BBa_K3407006). It corresponds to 27nt from the eGFP gene (nt 78 to 105), and is the one recognised and processed by Dicer to form siRNA. The shRNA has a single-stranded RNA (ssRNA) mutated loop region designed to deplete interaction with Fox-1 RBD (BBa_K3407004), containing its C3U and G6A mutations “UGUAUAU” in Fox-1 RBD's RNA target sequence. The dsRNA region contains a GG overhang that would represent the overhang left when mini-3 (BBa_K3407002) cleaves a tshRNA (See iGEM TU Delft 2020, Design - Future perspectives), and that represents a good substrate to process the shRNA by Dicer into a siRNA.