Difference between revisions of "Part:BBa K3407007:Design"
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===Design Notes===
 
===Design Notes===
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This biobrick represents the DNA template for the transcription of a 65 nt shRNA, excluding the promoter. Trancription was performed with <html><a id="6" href="https://parts.igem.org/Part:BBa_K3407023" target="_blank"><b>BBa_K3407023</b></a></html>. 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 (<html><a id="6" href="https://parts.igem.org/Part:BBa_K3407004" target="_blank"><b>BBa_K3407004</b></a></html>), containing its C3U and G6A mutations “<html><u>UG<b>U</b>AU<b>A</b>U</u><html>” in Fox-1 RBD's RNA target sequence. The dsRNA region contains a GG overhang that would represent the overhang left when mini-3 (<html><a id="6" href="https://parts.igem.org/Part:BBa_K3407002" target="_blank"><b>BBa_K3407002</b></a></html>) cleaves a tshRNA (See <html><a id="6" href="https://2020.igem.org/Team:TUDelft" target="_blank"><b>iGEM TU Delft 2020</b></a></html>, Design - Future perspectives), and that represents a good substrate to process the shRNA by Dicer into a siRNA.
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===Source===
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Synthetically synthesised
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===References===
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Revision as of 10:25, 27 October 2020


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.