Cell-Free Transcription-Translation: From Gene Circuits to Self-Assembly in a Test Tube

Cell-free transcription-translation (TX-TL) systems are becoming powerful platforms to construct complex biological systems in vitro through the expression of synthetic genetic circuits. In the past decade, considerable efforts have been made to expand the capabilities of those systems.

My lab has developed an all E. coli cell-free TX-TL system to expand the repertoire of regulatory parts to 100s of elements for the construction and execution of gene circuits outside living cells in either test tube reaction, microfluidics or liposomes. New metabolisms energize gene expression for 8-10 hours to reach enough protein synthesis and program the self-assembly of active biological structures. The phages T7 and phix174 are synthesized in cell-free TX-TL reactions from their genomes. Both phages serve as model systems to study the relationship information to self-assembly. Encapsulated inside cell-sized phospholipid liposomes, the cell-free TX-TL system is used to construct a minimal cell from the bottom-up. I will present this cell-free synthetic biology platform and our last experiments.