ArticleScientific reports2016
CRMAGE: CRISPR Optimized MAGE Recombineering.
Article in Scientific reports, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 96 papers.
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Who cites it
96 citing papers in PubMed, 227 citations in OpenAlex.
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- EASY-edit: a toolbox for high-throughput single-step custom genetic editing in bacteria.Nucleic acids research · 2025Article
- Genome-Editing Tools for Lactic Acid Bacteria: Past Achievements, Current Platforms, and Future Directions.International journal of molecular sciences · 2025Review
- SELECT: high-precision genome editing strategy via integration of CRISPR-Cas and DNA damage response for cross-species applications.Nucleic acids research · 2025Article
- A diverse single-stranded DNA-annealing protein library enables efficient genome editing across bacterial phyla.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Quadruplet codon decoding-based versatile genetic biocontainment system.Nucleic acids research · 2025Article
- Harnessing CRISPR interference to resensitize laboratory strains and clinical isolates to last resort antibiotics.Scientific reports · 2025Article
- Mutations in the essential outer membrane protein BamA contribute to Escherichia coli resistance to the antimicrobial peptide TAT-RasGAPThe Journal of biological chemistry · 2025Article
- ReaL-MGE is a tool for enhanced multiplex genome engineering and application to malonyl-CoA anabolism.Nature communications · 2024Article
36 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A bottleneck in metabolic engineering and systems biology approaches is the lack of efficient genome engineering technologies. Here, we combine CRISPR/Cas9 and λ Red recombineering based MAGE technology (CRMAGE) to create a highly efficient and fast method for genome engineering of Escherichia coli. Using CRMAGE, the recombineering efficiency was between 96.5% and 99.7% for gene recoding of three genomic targets, compared to between 0.68% and 5.4% using traditional recombineering. For modulation of protein synthesis (small insertion/RBS substitution) the efficiency was increased from 6% to 70%. CRMAGE can be multiplexed and enables introduction of at least two mutations in a single round of recombineering with similar efficiencies. PAM-independent loci were targeted using degenerate codons, thereby making it possible to modify any site in the genome. CRMAGE is based on two plasmids that are assembled by a USER-cloning approach enabling quick and cost efficient gRNA replacement. CRMAGE furthermore utilizes CRISPR/Cas9 for efficient plasmid curing, thereby enabling multiple engineering rounds per day. To facilitate the design process, a web-based tool was developed to predict both the λ Red oligos and the gRNAs. The CRMAGE platform enables highly efficient and fast genome editing and may open up promising prospective for automation of genome-scale engineering.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.