RNA

Part:BBa_K4286223:Design

Designed by: Bingru Feng   Group: iGEM22_SZU-China   (2022-09-27)


shRNA (RPMK1-2)-1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 17
    Illegal BglII site found at 38
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We have confirmed that this siRNA sequence has a good binding ability to Rhizoctonia solani. The intrinsic order of this sequence is sense RNAi fragment — loop — antisense RNAi fragment. This sequence was assembled in the pET28a (+) plasmid containing the IPTG-inducible phage T7 promoter and subsequently transferred into RNase-deficient E. coli HT115 (DE3). In our project, shRNA will be industrially produced by E. coli on a large scale, and shRNA will be purified and sprayed on rice fields to inhibit Rhizoctonia solani.


Source

We input the CDS sequence of RPMK1-2 in the professional shRNA design webpage, and the RNAi fragment is automatically designed by the program. Our examination confirmed that the sense portion of this sequence perfectly matches the gene encoding RPMK1-2 of Rhizoctonia solani isolate R98.


References

[1]Tiwari, I.M., Jesuraj, A., Kamboj, R. et al. Host Delivered RNAi, an efficient approach to increase rice resistance to sheath blight pathogen (Rhizoctonia solani). Sci Rep 7, 7521 (2017). https://doi.org/10.1038/s41598-017-07749-w