Optimizing the Receptor, aims for reduced background activation

In order to be able to implement a custom multiplexed transmembrane signal input/output relationship, the first condition is that engineering modular receptor to enable it to recognize extracellular signals and transduce them into customized intracellular signals. SynNotch is a transmembrane receptor with high programmability. Therefore, we can use this tool to receive extracellular signals and output customized intracellular signals.

Good tools are prerequisite to the successful execution of a job. In order to make SynNotch more suitable as a tool for receiving complex extracellular signals, we have optimized it. Please visit Optimization page for more details.

Three-layer design for dual transmembrane signals

The expression of membrane proteins on the cell membrane is limited. Our modeling tells us how to make cells sensitive to the limited signal molecules, and perform function efficiently is the key to make the transmembrane logic gate system work.

Based on this, we developed the ENABLE system with a 3-layers design: Receptor, Amplifier, and Combiner. By adding an intermediate layer, the “Amplifier” layer, we are able to effectively amplify the transmembrane signal, thus enabling our intracellular components to effectively travel logic functions.

The 3-layer design principle underlying ENABLE empowers any future development of transmembrane logic circuits, thus contributes a foundational advance to synthetic biology.

Unified intracellular logic gates layout

In order to be able to receive dual transmembrane signals and produce complete binary Boolean logic, we designed a set of interactive grammar for the interaction between the "Amplifier" layer and the "Combiner" layer within the cell membrane. This grammar consists mainly of the following elements, a transcription system based on the activating-form, silencing-from or NIMPLY-form promoters (the three are collectively referred to as a synthetic transcription factor-promoter pairs based transcription system); intein-based protein in vivo fusion systems, proteolytic enzyme-based protein in vivo destruction systems (the two are collectively referred to as a protein fusion/destruction-based transcription factor logic). We have test them and report in Results page, as well as on parts.igem pages.

ENABLE logic gates

Below we show experimental verifications of six (OR, NOR, AND, NAND, IMPLY, NIMPLY) of the eight truly binary logics.

Here, we use the OR gate as a proof of concept for a three-layer transmembrane binary Boolean logic during tissue patterning.

Through dynamic and static observations, we can see that the Receiver cell with transmembrane OR gate can be activated by two Sender cells expressing different membrane antigens.

Public engagement and education

Our human practice activities include dialogues (debate and interviews), presentations, Bio-Art display and hands-on practices, all of which centered around farseeing. We want to unlock the creativity and intelligence in as many people as possible. Fully engaged with the public, we motivated by their openness and willingness, which reminds us why we initiated the research and why we work hard in the lab - for the good of the people, including ourselves.