Composite

Part:BBa_K2412001:Design

Designed by: Jennifer Fang   Group: iGEM17_ColumbiaNYC   (2017-10-26)


shRNA circuit for eGFP knockdown


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
    COMPATIBLE WITH RFC[1000]


Design Notes

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


Source

This circuit uses the standard T7 promoter, and the shRNA was designed against the sequence of commercialized eGFP.

References

The shRNA sequence was published on https://www.addgene.org/30323/. 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.