Team:Fudan/Notebook

2018 iGEM Team:Fudan - Notebook

Our notebook

Here we present a main outline of our progress throughout the year.

Our notebook

Here we present a main outline of our progress throughout the year.

Monthly Highlights

April

  • First functional SynNotch constructed
  • Developed a simple testification method of SynNotch—shaking beads linked with antigen
  • Transcription factor and promotor pairs constructed and tested

May

  • SynNotch leakage problem detection
  • Successfully constructed and tested the first mutated version of SynNotch
  • Started constructing composite promoters for logic gates

June

  • SynNotch with various extra- and intra-cellular domain construction
  • SynNotch and composite promoter verification in cell

August

  • Batch construction of plasmids used in “amplifier” and “combiner” layers for all sixteen logic
  • Continual construction and testification of various mutation versions of SynNotch

September

  • Successfully constructed double-stable cell line of two SynNotch receptors
  • Finished constructing plasmids used in logic gates

October

  • Parts construction
  • All sixteen logic testification in cell
2018 team Fudan abstract

Abstract

Contact-dependent signaling is critical for multicellular biological events, yet customizing contact-dependent signal transduction between cells remains challenging. Here we have developed the ENABLE toolbox, a complete set of transmembrane binary logic gates. Each gate consists of 3 layers: Receptor, Amplifier, and Combiner. We first optimized synthetic Notch receptors to enable cells to respond to different signals across the membrane reliably. These signals, individually amplified intracellularly by transcription, are further combined for computing. Our engineered zinc finger-based transcription factors perform binary computation and output designed products. In summary, we have combined spatially different signals in mammalian cells, and revealed new potentials for biological oscillators, tissue engineering, cancer treatments, bio-computing, etc. ENABLE is a toolbox for constructing contact-dependent signaling networks in mammals. The 3-layer design principle underlying ENABLE empowers any future development of transmembrane logic circuits, thus contributes a foundational advance to Synthetic Biology.