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Title: Versatile seamless DNA vector production in E. coli using enhanced phage lambda integrase
Authors: Roy, Suki
Peter, Sabrina
Dröge, Peter
Keywords: Science::Biological sciences
Issue Date: 2022
Source: Roy, S., Peter, S. & Dröge, P. (2022). Versatile seamless DNA vector production in E. coli using enhanced phage lambda integrase. PLOS ONE, 17(9), e0270173-.
Project: NRF-CRP21-2018-0002
Journal: PLOS ONE
Abstract: Seamless DNA vectors derived from bacterial plasmids are devoid of bacterial genetic elements and represent attractive alternatives for biomedical applications including DNA vaccines. Larger scale production of seamless vectors employs engineered Escherichia coli strains in order to enable tightly regulated expression of site-specific DNA recombinases which precisely delete unwanted sequences from bacterial plasmids. As a novel component of a developing lambda integrase genome editing platform, we describe here strain MG1655-ISC as a means to easily produce different scales of seamless vectors, ranging in size from a few hundred base pairs to more than ten kilo base pairs. Since we employed an engineered lambda integrase that is able to efficiently recombine pairs of DNA crossover sites that differ in sequence, the resulting seamless vectors will be useful for subsequent genome editing in higher eukaryotes to accommodate variations in target site sequences. Future inclusion of single cognate sites for other genome targeting systems could enable modularity. These features, together with the demonstrated simplicity of in vivo seamless vector production, add to their utility in the biomedical space.
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0270173
Schools: School of Biological Sciences 
Rights: © 2022 Roy et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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