Part:BBa_K4830020:Design
TLR ngRNA 1
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The design of TLR ngRNA 1 includes the U6 promoter, spacer, gRNA scaffold and RTTPBS.
Source
The TLR pegRNA 1 design contains the spacer and RTTPBS sequence, which was adapted from other pegRNAs 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 132777 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.