Introduction
Synthetic minimal cells are new ways to experiment with living organism. They are engineered structures with proper cell partitions inside which we can perform many functions of a living cell. The advantage of this type of cells is that as if performs only one function independent of other processes in the cells, we can have independent studies uninterrupted by other cell processed.
Design an useful synthetic minimal cell
Design a system that can perform a function (biosensor, biomanufacturing, research - or any other purpose). Design all the steps of building and deploying the system: define function, pick components, describe results.
1. Pick a function.
1A. What would your synthetic cell do? What is the input and what is the output.
My synthetic minimal cell will act as a biosensor for lead ions. These cells will indicate the presence and
concentration on lead ions in an environment with bioluminance.
The input will be lead ions (Pb2+) and output will be light whose intensity will be proportional to the
concentration of lead icons.
1B. Could this function be realized by cell free Tx/Tl alone, without encapsulation?
No, i want to deploy the cells as biosensors in sewage and other places of lead pollution. Thus encapsulation is essential.
1C. Could this function be realized by genetically modified natural cell?
Yes, this can be realized by GM cells but with a synthetic minimal cell i am expecting a simple system without interference from other functions of the cell.
1D. Describe the desired outcome of your synthetic cell operation.
This cells will sense lead in sewage.
2. Design all components that would need to be part of your synthetic cell.
2A. What would be the membrane made of?
The cell need to have membrane permeable to lead ions. Another possible option would be to have lead specific receptors on the membrame. I would typically go for Phospholipid membrane.
2B. What would you encapsulate inside? Enzymes, small molecules.
The cell will have lead binding protein which will act as a promotor for bioluminance proteins. Other that that it will have the translation, transcription machinery's.
2C. Which organism your tx/tl system will come from? is bacterial OK, or do you need mammalian system for some reason? (hint: for example, if you want to use small molecule modulated promoters, like Tet-ON, you need mammalian system.
E.coli system will be good for this project.
2D. How will your synthetic cell communicate with the environment? (hints: are substrates permeable? or do you need to express membrane channel?
I would prefer a permeable membrane channel mechanism over expression of specific molecule uptake proteins, as it would be simple in construction and operation.
3) Experimental details
3A. List all lipids and genes (bonus: find the specific genes; for example, instead of just saying “small molecule membrane channel” pick actual gene)
LUX a, b genes, PBRr lead binding protein.
3B. How will you measure the function of your system?
Its a biosensor, thus the accuracy is predicting the lead concentration in comparison with a alternate sensing system would be an ideal test for the system.
Lab Task
Express GFP in cell free system
The shipping cost of reagents are too high we are not able to afford it. Trying to get them from alternate sources.
Ethics/ safety considerations this week
Do your activities this week raise new ethics and/or safety considerations you had not considered in week 1? Describe what activities have raised these considerations and any changes you have implemented in response.
No, as this is a design assignment so far.