RNA

Part:BBa_K2340002:Design

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


Guide RNA construct of human ꞵ-actin (2)


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 ꞵ-actin 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 ꞵ-actin 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.