Discussing the Science of Big DNA
In the latest issue of DECODED, the quarterly newsletter from Integrated DNA Technologies (IDT), Dr Dan Gibson discusses the science of big DNA. Dr Gibson spent most of the last decade working at the J Craig Venter Institute, as well as Synthetic Genomics Inc. (SGI), to develop, establish, and commercialize methods to take synthetic biology beyond its current limitations. In 2010 Dr Gibson and his team produced a DNA construct of 1.1 Mb, the largest synthetic DNA sequence to date, something which had not been achievable using traditional restriction/ligation methodologies. Instead, the team developed the Gibson Assembly™ Method, which uses a unique enzyme mix to assemble DNA elements with 20–80 overlapping bases. Once reamplified, the population, enriched with the correct sequence, is much faster to screen. The technology makes possible the creation of 10–30 kb sequences—the size of an entire biological pathway—in less than a week, and without cloning into E. coli. Dr Gibson and his team are currently looking at ways in which they can refine DNA assembly methods; improve large gene delivery to host cells; and develop methods that rely less on host organisms for copying their synthetic constructs.