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Programmable genome engineering
The model bacterium Escherichia coli has a single circular chromosome. Wang et al. created a method to fragment the E. coli genome into independent chromosomes that can be modified, rearranged, and recombined. The efficient fission of the unmodified E. coli genome into two defined, stable pairs of synthetic chromosomes provides common intermediates for large-scale genome manipulations such as inversion and translocation. Fusion of synthetic chromosomes from distinct cells generated a single genome in a target cell. Precise, rapid, large-scale genome engineering operations are useful tools for creating diverse synthetic genomes.
Science, this issue p. 922
Abstract
The design and creation of synthetic genomes provide a powerful approach to understanding and engineering biology. However, it is often limited by the paucity of methods for precise genome manipulation. Here, we demonstrate the programmed fission of the Escherichia coli genome into diverse pairs of synthetic chromosomes and the programmed fusion of synthetic chromosomes to generate genomes with user-defined inversions and translocations. We further combine genome fission, chromosome transplant, and chromosome fusion to assemble genomic regions from different strains into a single genome. Thus, we program the scarless assembly of new genomes with nucleotide precision, a key step in the convergent synthesis of genomes from diverse progenitors. This work provides a set of precise, rapid, large-scale (megabase) genome-engineering operations for creating diverse synthetic genomes.
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