RNA

Part:BBa_K2340007:Design

Designed by: Laurens Heling   Group: iGEM17_Kent   (2017-10-31)


Guide RNA construct of human Pkp4 (1)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 217
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 183
    Illegal XhoI site found at 224
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence complementary to the human Pkp4 had to be exclusively complementary to that. If complementary to other mRNA sequences, the Cas13 could target those as well


Source

Sequences for crRNA backbone were taken from East-Selesky et al (2016).


The complementary sequence to human Pkp4 mRNA and the sequences of the human U6 promoter and terminator are taken from Genbank


References

East-Selesky, A. et al. (2016) Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature 538: 270–273.