Difference between revisions of "Part:BBa K3738027:Design"

 
(→‎References)
 
Line 16: Line 16:
  
 
===References===
 
===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-cas1

Latest revision as of 03:53, 22 October 2021


Lbu-Cas13a with an N-terminal Anionic Tag and C-Terminal 6xHistidine Tag and crRNA


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 443
    Illegal BglII site found at 1379
    Illegal BglII site found at 1589
    Illegal BglII site found at 1643
    Illegal BglII site found at 1874
    Illegal BglII site found at 2147
    Illegal BglII site found at 2633
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1162
    Illegal PstI site found at 2017
    Illegal PstI site found at 2956
    Illegal NgoMIV site found at 2095
    Illegal NgoMIV site found at 2731
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part has been codon optimized for E.coli. 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. The Cas13a protein comes from the Gram-negative bacteria Leptotrichia buccalis (Lbu).

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-cas1