Part:BBa_K5131004

FlipGFP_nsp5 inducible(10-11)

FlipGFP is a fluorescent protein dependent on protease activation[1]. In FlipGFP, a dimerization linker (E5 and K5) is inserted between β10 and β11, causing them to align parallel to each other and thus preventing fluorescence by failing to self-assemble with β1-9. By inserting a specific protease recognition sequence between β11 and K5, the protease can recognize and cleave this sequence(Figure 1). Subsequently, β10 and β11 can rearrange into their normal antiparallel positions, restoring fluorescence. This design provides FlipGFP with high specificity and flexibility for detection. By simply replacing the linker between β11 and K5, FlipGFP can be tailored to respond to different proteases.

Figure 1. Activation mechanism of FlipGFP[1]

This part includes the 10-11 β-strands of FlipGFP where the protease recognition sequence linked before β11 was replaced with the nsp5 recognition sequence(N-AVLQSGFRK-C).

Reference:
1. Zhang Q, Schepis A, Huang H, Yang J, Ma W, Torra J, Zhang SQ, Yang L, Wu H, Nonell S, Dong Z, Kornberg TB, Coughlin SR, Shu X. Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals. J Am Chem Soc. 2019 Mar 20;141(11):4526-4530. doi: 10.1021/jacs.8b13042. Epub 2019 Mar 6. PMID: 30821975; PMCID: PMC6486793.

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]