Pier Luigi Luisi Synthetic Biology Lab
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The Minimal Cell

The general goals of this research is the construction of vesicle-based bioreactors that simulate the main functions of biological cells.
The main technique here is the introduction of genes and enzymes into vesicles.
The final goal is the construction of a vesicular system that contain the minimal and sufficient number of genes and other components to be defined as living (self-sustaining + self-reproduction + evolvability).

The main researchers in this filed are Pasquale Stano and Giovanni Murtas, as well as Yutetsu Kumura from the Tokyo University (from the group of prof. Ueda, with whom we are collaborating).

Supporting Projects on Liposomes and other Compartments

Fusion of Vesicles:
the real fusion of vesicles with different content in the water pool is the basis for the construction of vesicles having a high degree of internal complexity. This is studied using vesicles oppositely charged (e.g. oleate vs. DDAB vesicles), with all needed physical chemistry (phase diagrams and check of the fusion efficiency by fluorescence techniques).
Filippo Caschera is presently working in the area.

Characterization of Phosphatidic Acid Liposomes:
phosphatidic acid is the biochemical intermediate to the biosynthesis of lecithin, and in one of our projects for the minimal cells was important to see whether the corresponding liposomes are endowed with sufficient stability and capability as bioreactors. Julien Dubois has been working with this project.

Study of “Bola” Bicarboxylic Acid Systems as Possible Prebiotic Membrane-Forming Compounds:
in the group of Sandra Pizzarello, at the Arizona State University, with whom we collaborate, long chain a-w dicarboxylic acids have been found in meteorites-and therefore can be considered as prebiotic surfactants.

The study of their capability of forming aggregates is in progress, Chris Thomas is presently studying this together with the student Armando Romani.

Interaction of Vesicles with RNA:
Chris Thomas as part of his work for the doctorate has been studying the interaction of t-RNA with vesicles, to see whether RNA is capable of discriminating the vesicle size.

The Use of Water-in-oil Emulsion as Bioreactors:
water in oil microemulsion (reverse micelles) and emulsion have the advantage of offering compartments with compulsory affinity for water-soluble substrates.
Deborah Fiordemondo in her thesis has been studying the expression of the green fluorescent protein by mixing w/o emulsion droplets containing the various molecular biology components.



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