Genome-scale genetic engineering in Escherichia coli

Recent advances and versatility of MAGE towards industrial applications

“…However, MAGE represents a versatile and powerful toolkit because of its simplicity, rapid experimental practice and highly efficient genome engineering capability (Jeong et al 2013). …”

“…Metabolic engineering has the potential to produce high yields of desired products in a cost effective manner by the extension of pathways or editing of genomes towards industrial scale at competitive prices. MAGE shows a rapid, specific and efficient tool for generating billions of genome diversity that can be useful for high level production and optimization of biosynthetic pathways Jeong et al 2013;Isaacs et al 2011;Lajoie et al 2013) and this technique has dramatically increased our ability to engineer cells in a directed and combinatorial manner towards the development of flexibly programmable cells (Carr et al 2012;Esvelt and Wang 2013).…”

Rapid and high-throughput construction of microbial cell-factories with regulatory noncoding RNAs

“…However, since it still lacks well-established engineering design principles for identifying effective target genes, metabolic engineering must be carried out through trial and error. Genome editing is also limited to gene insertion or deletion at random or at a few pre-defined locations across the genome (Esvelt and Wang, 2013;Jeong et al, 2013;Song et al, 2015). Moreover, technical implementation of this editing takes a week or more, which is long compared to other conventional processes and accordingly makes genome editing laborious and expensive (Esvelt and Wang, 2013;Jeong et al, 2013;Song et al, 2015).…”

“…Genome editing is also limited to gene insertion or deletion at random or at a few pre-defined locations across the genome (Esvelt and Wang, 2013;Jeong et al, 2013;Song et al, 2015). Moreover, technical implementation of this editing takes a week or more, which is long compared to other conventional processes and accordingly makes genome editing laborious and expensive (Esvelt and Wang, 2013;Jeong et al, 2013;Song et al, 2015). In addition to the identification of effective target genes, utilization of an appropriate bacterial strain that possesses inherently higher production capability is another issue in metabolic engineering.…”

Engineering

“…Likewise, the increased use of combinatorial and evolutionary approaches, fueled by a rapid expansion of synthetic molecular tools, opened the possibility for testing novel and large combinations of gene expression systems and genetic backgrounds [14,15]. Additionally, efforts concerning the optimization of fermentation conditions have succeeded in scaling-up many AAA production processes, while simultaneously providing important feedback on the physiological behavior of engineered strains [16,17].…”