RNA

Part:BBa_K2624004:Design

Designed by: Shuyao Zhou   Group: iGEM18_CPU_CHINA   (2018-08-26)


U6-inhSi-pri-miRNA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 312
    Illegal NgoMIV site found at 341
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 305


Design Notes

Please refer to the wiki of CPU_CHINA 2018 because this part is strictly related to the other parts used in the project.
The sequence of the inhibitory strand is dependent on the pri-miRNA basal segments that we target. Then we need to make sure that the actual appearance of the product of this part is indeed only the wanted sequence, with no 'tail'. So we added a hammerhead ribozyme in the 3'-end, and a RNA pol III promoter.

Source

The U6 promoter is from BBa_K2200003 and it is widely used, inside or outside the iGEM competition. The inhibitory strand is purly through our design. We did not refer to any public database.

References

[1]Dahlberg J E, Lund E. The genes and transcription of the major small nuclear RNAs[M]//Structure and function of major and minor small nuclear ribonucleoprotein particles. Springer, Berlin, Heidelberg, 1988: 38-70.
[2]Kao C C, Yang X, Kline A, et al. Template requirements for RNA synthesis by a recombinant hepatitis C virus RNA-dependent RNA polymerase[J]. Journal of virology, 2000, 74(23): 11121-11128.
[3]Han J, Lee Y, Yeom K H, et al. Molecular basis for the recognition of primary microRNAs by the Drosha-DGCR8 complex[J]. Cell, 2006, 125(5): 887-901.