Journal
BIOENGINEERED
Volume 6, Issue 3, Pages 136-140Publisher
TAYLOR & FRANCIS INC
DOI: 10.1080/21655979.2015.1011029
Keywords
directed evolution; DNA assembly; enzyme; genome engineering; metabolic engineering; recombineering; synthetic biology
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Funding
- National Natural Science Foundation of China [31200020]
- Major State Basic Research Development Program of China (973 Program) [2014CB745103]
- Natural Science Foundation of Jiangsu Province [BK20141107]
- Key Technologies R & D Program of Jiangsu Province, China [BE2014607]
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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.
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