4.3 Review

Recombineering and MAGE

Journal

NATURE REVIEWS METHODS PRIMERS
Volume 1, Issue 1, Pages -

Publisher

SPRINGERNATURE
DOI: 10.1038/s43586-020-00006-x

Keywords

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Funding

  1. US Department of Energy (DOE) [DE-FG02-02ER63445]
  2. National Institute of General Medical Sciences of the National Institutes of Health (NIH) under a Chemistry-Biology Interface Training Grant [T32GM133395]
  3. European Research Council (ERC) [H2020-ERC-2014-CoG 648364]
  4. Hungarian Academy of Sciences [LP-2017-10/2017, KKP 126506, GINOP-2.3.2-15-2016-00014]
  5. Physical and Engineering Biology training grant [5T32EB019941-05]
  6. EMBO [LTF 160-2019]
  7. Air Force Office of Scientific Research [FA9550-14-1-0089]
  8. Welch Foundation [F-1654]

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Recombineering and MAGE have revolutionized genetic engineering, offering powerful new methods to simplify genetic manipulation and improve throughput in experimental biology. These tools have the potential to transcend boundaries between prokaryotes and eukaryotic cells.
Recombination-mediated genetic engineering, also known as recombineering, is the genomic incorporation of homologous single-stranded or double-stranded DNA into bacterial genomes. Recombineering and its derivative methods have radically improved genome engineering capabilities, perhaps none more so than multiplex automated genome engineering (MAGE). MAGE is representative of a set of highly multiplexed single-stranded DNA-mediated technologies. First described in Escherichia coli, both MAGE and recombineering are being rapidly translated into diverse prokaryotes and even into eukaryotic cells. Together, this modern set of tools offers the promise of radically improving the scope and throughput of experimental biology by providing powerful new methods to ease the genetic manipulation of model and non-model organisms. In this Primer, we describe recombineering and MAGE, their optimal use, their diverse applications and methods for pairing them with other genetic editing tools. We then look forward to the future of genetic engineering. This Primer by Wannier and colleagues summarizes the methodology, analysis and utility of recombineering and multiplex automated genome engineering (MAGE) in microbial species. In addition, this Primer examines advanced techniques that pair MAGE with other tools to improve editing efficiency.

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