Composite

Part:BBa_K3738025:Design

Designed by: Mark Lea   Group: iGEM21_Lethbridge   (2021-10-20)
Revision as of 17:21, 20 October 2021 by Marklea (Talk | contribs) (→‎References)

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McyH complementary Lbu-crRNA (Composite)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This composite part includes the IPTG-inducible T7 promoter (BBa_J64997), McyH Complementary Lbu-crRNA (BBa_K3738023), and double terminator BBa_B0015.

In design of crRNA, ensure the final complementary base is not a G.

The part must be in vitro transcribed to obtain the direct-repeat stem loop RNA structure and complex with Cas13a.


Source

The Mcy gene cluster comes from Microcystic Aeruginosa, however the crRNA originates from Leptrotrichia buccalis

References

Glasgow, J., Capehart, S., Francis, M., and Tullman, D (2012) Osmolyte-Mediated Encapsulation of Proteins inside MS2 Viral Capsids. ACS Nano. 10, 8658-8664.

Huynh, N., Depner, N., Larson, R. et al. A versatile toolkit for CRISPR-Cas13-based RNA manipulation in Drosophila. Genome Biol 21, 279 (2020). https://doi.org/10.1186/s13059-020-02193-y

McDade Joel. CRISPR 101: Targeting RNA with Cas13a (C2c2). Retrieved from https://blog.addgene.org/crispr-101-targeting-rna-with-cas13a-c2c2.