Coding

Part:BBa_K3168001:Design

Designed by: Eva Hanckmann, Harm van der Veer, Claire Michielsen   Group: iGEM19_TU_Eindhoven   (2019-09-12)
Revision as of 12:47, 17 September 2019 by CMichielsen (Talk | contribs) (→‎Source)


dCas9-CP1041


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2143
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 261
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Designed for bacterial expression.

Source

Mutated variant of S. pyogenes derived Cas9, which was synthesised in gBlocks by IDT. The sequence was based on Addgene plasmid #119808.

References

Oakes, B. L., Fellmann, C., Rishi, H., Taylor, K. L., Ren, S. M., Nadler, D. C., ... & Savage, D. F. (2019). CRISPR-Cas9 circular permutants as programmable scaffolds for genome modification. Cell, 176(1-2), 254-267.

Park, J. J., Dempewolf, E., Zhang, W., & Wang, Z. Y. (2017). RNA-guided transcriptional activation via CRISPR/dCas9 mimics overexpression phenotypes in Arabidopsis. PloS one, 12(6), e0179410.

Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott, D. A., & Zhang, F. (2013). Genome engineering using the CRISPR-Cas9 system. Nature protocols, 8(11), 2281.

Sternberg, S. H., LaFrance, B., Kaplan, M., & Doudna, J. A. (2015). Conformational control of DNA target cleavage by CRISPR–Cas9. Nature, 527(7576), 110.