RNA

Part:BBa_K4830024:Design

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


PETLR ngRNA 1


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 PETLR ngRNA 1 includes the U6 promoter, spacer and gRNA scaffold.

Source

The PETLR ngRNA 1 design contains the spacer, 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 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.