Project

Part:BBa_K3218005

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


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

This is the second mutation mechanism that was designed and it consists of a 'dead' deactivated Cas9 which serves as the binding domain and it is fused to an eAID protein which induces mutations through the chemical change of a cytosine to a uracil. Unlike the dCas9-FokI (BBa_K3218003) this mechanism only needs one gRNA and therefore only one intron and can be widely spaced and it does not cause a double stranded break. The eAID is different than AID in that is has a larger range for its mutating mechanism.

Sequence and Features


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


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