Difference between revisions of "Part:BBa K4830023:Design"

 
 
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===Design Notes===
 
===Design Notes===
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The design of TLR pegRNA 2 includes the U6 promoter, spacer, gRNA scaffold and RTTPBS.
+
The design of PETLR pegRNA 1 includes the U6 promoter, spacer, gRNA scaffold and RTTPBS.
−
 
+
−
 
+
  
 
===Source===
 
===Source===
 +
The PETLR 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.
  
−
The TLR pegRNA 2 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.
  
−
===References===
+
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.

Latest revision as of 15:40, 11 October 2023


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

Source

The PETLR 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.