This invention is a reconfigurable chassis that allows for rapid construction and optimization of biocircuits.
Technologies for cloning and recombination of genetic material are time-consuming and rate-limiting. For instance, transformed plasmids are less stable than chromosomal DNA and are limited in the number of different plasmids with which they can be co-transformed. This invention is a reconfigurable chassis enabling rapid construction of biocircuits.
The BioFPGA chassis, named for Field-Programmable Gate Arrays (FPGAs), is a cellular platform that supports reprogrammable logic. A BioFPGA allows rapid prototyping of new regulatory networks by providing specific structures and scaffolding ahead of time and does not require DNA assembly or plasmid design, which may be time-consuming, tedious and error-prone. Since no new plasmids are incorporated, this method is simpler, safer, and may be done by less experienced users. An unprogrammed or "blank" BioFPGA consists of a chromosomal sequence of alternating att-sites and counterselection markers. First, allelic replacement alters the recombination sites to specify which part from a library will go into which target site. Then the library parts are integrated in the chromosome at the user-specified locations. The library may contain any useful parts even if constructed by other means. This technology allows a designer to apply a MAGE-like protocol (Church), followed by a recombination step, to a predesigned configurable chassis, to support rapid prototyping of new biocircuits.
- No DNA assembly or plasmid design required
may include parts constructed by other means; All possible combinations of
library part to target chromosomal locations are available
quick, simple and safe programming of biocircuits by less sophisticated users