Part:BBa_K3431001

pp21_B Toehold Switch for miR-21 Detection

This toehold switch has been designed to open up its hairpin loop structure upon binding with miRNA-21, resulting in The translation of downstream reporter protein. The design of toehold switch can be separated into the following 7 region from its 5' end: trigger binding sites (with extended base pairs upfront to improve the stability of toehold switch mRNA secondary structures), stem region, loop region with RBS, complimentary stem region, start codon, complimentary stem region, and linker amino acids. For this particular toehold switch (pp21), we derived its design based on an articles indicating novel designs of toehold switches for detecting miRNA-21 and miRNA-155 in HEK-293 cells (Wang, S et.al). We incorporate the loop and linker sequence from the studies to test out whether optimisations specific to mammalian cells can have a better regulatory function for downstream protein expression in PURExpress in vitro protein synthesis system, on which our project of diagnosing oral cancer with toehold switch and glucose meter are based.

References: Wang, S., Emery, N. J., & Liu, A. P. (2019). A novel synthetic toehold switch for microRNA detection in mammalian cells. ACS synthetic biology, 8(5), 1079-1088.

Sequence and Features

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]