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Part:BBa_K3218004:Design

Designed by: Abraham Grosman   Group: iGEM19_MSP-Maastricht   (2019-10-15)


CRISPR-X: deactivated Cas9 coupled to activation induced deaminase (AID)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5456
    Illegal NheI site found at 6011
    Illegal NotI site found at 1146
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3001
    Illegal BglII site found at 3460
    Illegal BglII site found at 3549
    Illegal BglII site found at 4194
    Illegal BamHI site found at 669
    Illegal XhoI site found at 5989
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 286
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1236
    Illegal SapI site found at 1337


Design Notes

As said before for this mechanism only one intro was required, but careful design is still required since the orientation of the intron/gRNA decides on which side of the intron the AID will be mutating.


Source

the dCas9 comes from a bacterial genome and the mutating mechanism is human AID

References