title: Directed evolution combined with synthetic biology strategies expedite semi-rational engineering of genes and genomes.
abstract: Owing to our limited understanding of the relationship between sequence and function and the interaction between intracellular pathways and regulatory systems, the rational design of enzyme-coding genes and de novo assembly of a brand-new artificial genome for a desired functionality or phenotype are difficult to achieve. As an alternative approach, directed evolution has been widely used to engineer genomes and enzyme-coding genes. In particular, significant developments toward DNA synthesis, DNA assembly (in vitro or in vivo), recombination-mediated genetic engineering, and high-throughput screening techniques in the field of synthetic biology have been matured and widely adopted, enabling rapid semi-rational genome engineering to generate variants with desired properties. In this commentary, these novel tools and their corresponding applications in the directed evolution of genomes and enzymes are discussed. Moreover, the strategies for genome engineering and rapid in vitro enzyme evolution are also proposed.
authors.affiliation: a Key Laboratory of Industrial Biotechnology; Ministry of Education ; Jiangnan University ; Wuxi , Jiangsu China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Kang
authors.forename: Junli
authors.initials: J
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Jin
authors.forename: Sen
authors.initials: S
authors.lastname: Yang
mesh_terms: D004247:DNA / Q000235:genetics*; D053938:DNA Fragmentation; D019020:Directed Molecular Evolution; D016678:Genome; D057166:High-Throughput Screening Assays; D010641:Phenotype; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D004247:DNA
keywords: DNA assembly; HTS, high-throughput screening; LCR, Ligase Cycling Reaction; MAGE, multiplex automated genome engineering; directed evolution; dsDNA, double-stranded DNA; enzyme; genome engineering; metabolic engineering; recombineering; ssDNA, single-stranded DNA; synthetic biology