RNA

Part:BBa_K4830026:Design

Designed by: Ong Jie Hao   Group: iGEM23_NTU-Singapore   (2023-10-11)


777BFP ngRNA


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

The 777BFP ngRNA includes the spacer and gRNA scaffold.


Source

The 777BFP ngRNA design contains the spacer sequence, which was adapted from other ngRNAs in the Prime Editor 2 paper cited in References. With the help of TWIST Bioscience we synthesized those and other necessary sequences, and further combined with 65777 with BFP insert backbone in our lab.

References

Anzalone AV, Randolph PB, Davis JR, Sousa AA, Koblan LW, Levy JM, et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature. 2019 Oct 21;576.

Certo, M. T., Ryu, B. Y., Annis, J. E., Garibov, M., Jarjour, J., Rawlings, D. J., & Scharenberg, A. M. (2011). Tracking genome engineering outcome at individual DNA breakpoints. Nature Methods, 8(8), 671–676.