RNA

Part:BBa_K4286909:Design

Designed by: Ruoyu Chen，Minxi Zeng   Group: iGEM22_SZU-China   (2022-09-11)

shRNA molecules that can silence the gene encoding Ubiquitin ligase E3.


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]


Assembly

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 PG 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 PG of Rhizoctonia solani AG1-IA.


References

[1]Zhou B, Bailey A, Niblett CL, Qu R. Control of brown patch (Rhizoctonia solani) in tall fescue (Festuca arundinacea Schreb.) by host induced gene silencing. Plant Cell Rep. 2016 Apr;35(4):791-802. doi: 10.1007/s00299-015-1921-7. Epub 2016 Jan 22. PMID: 26800976. [2]Bilir Ö, Göl D, Hong Y, McDowell JM, Tör M. Small RNA-based plant protection against diseases. Front Plant Sci. 2022 Aug 18;13:951097. doi: 10.3389/fpls.2022.951097. PMID: 36061762; PMCID: PMC9434005. [3]Rabuma T, Gupta OP, Chhokar V. Recent advances and potential applications of cross-kingdom movement of miRNAs in modulating plant's disease response. RNA Biol. 2022;19(1):519-532. doi: 10.1080/15476286.2022.2062172. Epub 2021 Dec 31. PMID: 35442163; PMCID: PMC9037536.