Translational_Unit

Part:BBa_K2615003

Designed by: Yunqian Zhang   Group: iGEM18_OUC-China   (2018-08-17)
Revision as of 02:04, 18 August 2018 by Registry (Talk | contribs)

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Csy4-WT, the No.1 member of Csy4 family.

Csy4, one of Cas6 endoribonucleases, recognizes a specific 28-nt RNA and cleaves between 20G and 21C. It is possible to cleave an engineered Csy4 site in the coding region of a gene repression system. In our project, we constructed a Cas-based translational activator system. This cleavage releases a cis-repressive RNA module (crRNA) from the masked ribosome binding site (RBS), which subsequently allows the downstream translation initiation. Unlike small RNA as a translational activator, the endoribonuclease-based activator is able to efficiently unfold the perfect RBS-crRNA pairing. We found that, upon Csy4 induction, the new crRNA-RBS duplex can dramatically increase the GFP intensity, reaching the maximallevel of GFP in SC control. As a result, compared with the control group，the GFP fluorescence can be increased ***fold by Csy4, indicating that adjusting the strength of the RBS-crRNA duplex is an effective route to optimize translation activation of cis-repressed sfgfp mRNA.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 377
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 93


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Categories
Parameters
None