The combination of synthetic gene networks with in vitro protein production technology opens new innovative fields of application in medicine and biotechnology. Examples are environmental sensors, controls for biomanufacturing of biofuels, stem cell medicine, gene therapy (CRISPR), and manufacturing of functional materials. Cell-free platforms help to overcome limitations occurring in living host organisms, which are typically tied to the biochemical cross-talk between the host cell and the artificial gene circuit and a consequence of the potentially cytotoxic effect of the exogeneous gene network on the living cell on the one hand, and the availability of resources for transcription and translation, DNA replication and metabolites on the other. As those resources are dependent on the cell density, growth rate and cultivation conditions, deploying cell-free systems for gene circuit testing creates a more controlled environment thus increasing reproducibility and robustness of the circuit behavior and output.
NOTE: The Toolbox 2.0 Plasmid that you receive upon ordering is intended for propagation purposes only. To make the plasmid suitable for in vitro expression in myTXTL, please follow our recommended 2-step procedure to generate high-purity plasmid samples. For propagation of P70a-vector, please also read our Tech note about “Preparation of chemo-competent KL740 cells for amplification of P70a vectors” (available under Supplemental Material below).
The myTXTL® Toolbox 2.0 Plasmid Collection was designed and intensively studied by Vincent Noireaux, PhD at the University of Minnesota and co-workers. It contains over one hundred plasmids with various promoters and open reading frames (ORFs) to investigate gene regulation and molecule turnover. Available open reading frames include a wide selection of transcription factors, TXTL modulators and fluorescent reporter proteins to build multi-stage gene circuits.
A gene circuit executed in any myTXTL® kit is required to start with a σ70-specific promoter, as gene expression in myTXTL® relies entirely on the endogenous TXTL machinery of E. coli. Its modular and pre-designed format makes it also ideal for educational purposes to train students and demonstrate basic concepts of synthetic biology.