RNA

Part:BBa_K4830022:Design

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


TLR ngRNA 2


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 design of TLR ngRNA 1 includes the U6 promoter, spacer and gRNA scaffold.

Source

The TLR ngRNA 2 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 a 65777 backbone with a mCherry-stop-Clover insertion 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.