Part:BBa_K4375016

anti-EGFR Nanobody Display System

This construction consists of 4 described parts. These are: J23101* (BBa_K4375003), anti-EGFR-specific Nanobody: 7D12 (BBa_K4375005), Intimin (BBa_K4375012), Myc (BBa_K4375014). They allow the bacterium to bind to the cancer cell surface, thus allowing its visualization and colonization on the tumor surface.

Usage and Biology

Figure 1: Structure of the displayed construct. The image was created with BioRender.

This construct is meant for detecting cancer cells expressing EGFR receptors. It is expressed by the bacterial cells on their surface and can be used to detect the tumor by as it causes the bacteria to bind to cancer cells. This device contains two different parts whose production is controlled by a constitutive strong promoter J23101* (BBa_K4375003). The anti-EGFR Nanobody (BBa_K4375005) is at the N-terminal, tagged with a c-myc for detection. The Nanobody is fused to the Intimin part, which is a 12-stranded β-barrel protein found in bacterial adhesins (BBa_K4375012). Specific interaction can be detected by two tags: Myc (BBa_K4375014) and anti-E, or via the fluorescence of the sfGFP at the C-terminal.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 29
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 2029
  Illegal BamHI site found at 4045
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2050
  Illegal NgoMIV site found at 4066
  Illegal AgeI site found at 4124
  Illegal AgeI site found at 4256
  Illegal AgeI site found at 4640
  
• 1000
  COMPATIBLE WITH RFC[1000]

Sources

Piñero-Lambea, C.; Bodelón, G.; Fernández-Periáñez, R.; Cuesta, A. M.; Álvarez-Vallina, L.; Fernández, L. Á. Programming Controlled Adhesion of E. Coli to Target Surfaces, Cells, and Tumors with Synthetic Adhesins. ACS Synth. Biol. 2014, 4, 463–473. https://doi.org/10.1021/sb500252a.

Lim, B.; Yin, Y.; Ye, H.; Cui, Z.; Papachristodoulou, A.; Huang, W. E. Reprogramming Synthetic Cells for Targeted Cancer Therapy. ACS Synth. Biol. 2022, 11, 1349–1360. https://doi.org/10.1021/acssynbio.1c00631.