Composite

Part:BBa_K2412003:Design

Designed by: Jennifer Fang   Group: iGEM17_ColumbiaNYC   (2017-10-27)


circuit for shRNA knockdown of EGFR


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 43
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 89


Design Notes

The hairpin structure of the part made cloning challenging, but but assembling the part from two pieces with USER cloning made the assembly successful.


Source

The shRNA was designed against EGFR in human cancer cells.

References

The shRNA sequence was derived from a paper (the exact paper will be specified in a few days). The circuit design was published in Xiang, Shuanglin, et al. “Short hairpin RNA–expressing bacteria elicit RNA interference in mammals.” Nature Biotechnology, vol. 24, 14 May 2006, pp.697–702.