SynNotch receptors set

We have provided 7 versions of SynNotch receptors. Each of them can transduce signal from out of the cell via a contact-dependent antigen-stimulated cleavage process. When expressing two SynNotch receptors with two different extracellular domains recognizing two types of antigens, the cell is able to accept dual inputs. The chosen nanobodies are highly specific against their antigens, and the chosen intracellular domains are transcriptionally orthogonal. For more specific details, please refer to our parts listed below.

Transcriptional amplifiers set

Last year, we have constructed 3 zinc finger-based transcription repressors. This year, we have expanded our zinc finger-based transcription factors library by adding 3 more zinc finger-based transcription activators.

Although we prefer to use eight copies of response elements (RE) for a balance between not-too-hard molecular cloning and sufficient signal-to-noise ratio, we have provided 4 promotors with different repeats of response elements, allowing others to explore and tune their own transcriptional amplifiers. Latest modeling strongly supports our experimental preference, showing that eight copies of response elements can render the best result in line with our needs.

As is stated above, we have provided three types of transcriptional-based Amplifiers, including zinc finger-based, TEV protease-based and split intein-based. These Amplifiers must be used with our Combiners to execute designed binary logic function.

Amplifiers and Combiners work together to execute binary computation

Zinc finger-based

For simple binary logic function, such as A gate, NOT A gate, OR gate, NOR gate, and NIMPLY gate, placing response elements upstream and downstream of the promoter is sufficient. For more details, please visit our results page.

TEV protease-based

For IMPLY gate, we placed TEV protease-controlled destroyable nuclear localization sequence between zinc finger DNA binding domain (DBD) and transcription factor (TF). In the presence of intracellular TEV protease, DBD-TF would be cleaved, thus destroying its transcriptional regulation function and efficiently blocking its signal. For more details, please visit our results page.

Split intein-based

We built the more complex binary logic gates, such as XOR and XNOR gate, using split intein-based Amplifiers. Utilizing intein's ability to fuse the split ends of a transcription factor together, we're able to present these logic gates consistent with our 3-layer design principle. For more details, please visit our results page.

ENABLE - 16 logic gates for transmembrane signaling

Here is our 3-layer design of transmembrane binary logic gates. On the left lying a truth table which indicates the inputs and outputs of the according logic computation. What is set on the right is how it works. The double crossline on the top represents the plasma membrane. A and B signify the signal input, which are surface-expressed EGFP and surface-expressed CD19 in our system. What are embed on the membrane are our SynNotch receptors, the intracellular domain of which are transcriptional activators, which are tTAA and GV2 in our assay. The large rectangle represents the nucleus. The elongated rectangle with an array before represents the genetic circuit that are under control of the transcriptional factors. Squares in groups of three represent transcriptional factors that are amplified by the amplifier layer. You can swipe the bar to detect other gates by yourself.