ArticlePloS one2016
PCR-Based Seamless Genome Editing with High Efficiency and Fidelity in Escherichia coli.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Microbiota-derived succinate promotes enterohaemorrhagic Escherichia coli virulence via lysine succinylation.Nature microbiology · 2025Article
- A novel C-terminal degron identified in bacterial aldehyde decarbonylases using directed evolution.Biotechnology for biofuels · 2020Article
- Effective production of succinic acid from coconut water (Biotechnology reports (Amsterdam, Netherlands) · 2019Article
- Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing.Molecular biotechnology · 2018Article
- Construction of pyruvate producing strain with intact pyruvate dehydrogenase and genome-wide transcription analysis.World journal of microbiology & biotechnology · 2017Article
- High production of fatty alcohols in Escherichia coli with fatty acid starvation.Microbial cell factories · 2016Article
Corrections and comments
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Authors and funding
11 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Efficiency and fidelity are the key obstacles for genome editing toolboxes. In the present study, a PCR-based tandem repeat assisted genome editing (TRAGE) method with high efficiency and fidelity was developed. The design of TRAGE is based on the mechanism of repair of spontaneous double-strand breakage (DSB) via replication fork reactivation. First, cat-sacB cassette flanked by tandem repeat sequence was integrated into target site in chromosome assisted by Red enzymes. Then, for the excision of the cat-sacB cassette, only subculturing is needed. The developed method was successfully applied for seamlessly deleting, substituting and inserting targeted genes using PCR products. The effects of different manipulations including sucrose addition time, subculture times in LB with sucrose and stages of inoculation on the efficiency were investigated. With our recommended procedure, seamless excision of cat-sacB cassette can be realized in 48 h efficiently. We believe that the developed method has great potential for seamless genome editing in E. coli.
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Registered trials
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