RNA

Part:BBa_K1633007:Design

Designed by: Chen Xi, Zhou Yu, Jiang Waner, Zhang Peng, Tian Chenfei   Group: iGEM15_NJU-China   (2015-09-13)
Revision as of 14:52, 18 September 2015 by ChenXi (Talk | contribs) (→‎Design Notes)

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GFP siRNA


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]


Design Notes

We designed specific GFP siRNA based on a free software accessible online. This tool can find the best siRNA sequences on target gene GFP to insure the maximum gene-specificity and silencing efficacy. This tool also designs the pair of oligonucleotides needed to generate short hairpin RNAs (shRNAs) in the plasmid.

This part is artificial designed to target and downregulate GFP protein. This part is a short hairpin RNA (shRNA) sequence. When this shRNA sequence is cut by restriction enzyme and then integrated into pcDNA 6.2 vector, this shRNA can play a RNAi function in mammalian cell lines such as HEK293 cell. When the shRNA vector of GFP is transfected into mammalian cells, the shRNA hairpin structure is cleaved by Dicer into siRNA of GFP and loaded into the RISC. The siRNA-RISC complex targets at GFP mRNA under the guide of siRNA sequence and cleave the GFP mRNA.

Source

We designed the GFP 122 siRNA and ordered the part from a DNA synthesis company.


References