ArticlePloS one2011
OmniChange: the sequence independent method for simultaneous site-saturation of five codons.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- LCI Engineering for Improved Polystyrene Binding: The Impact of Aromatic Amino Acid Substitutions.ACS omega · 2025Article
- Synthetic directed evolution for targeted engineering of plant traits.Frontiers in plant science · 2024Review
- A primer to directed evolution: current methodologies and future directions.RSC chemical biology · 2023Review
- Thermostability engineering of industrial enzymes through structure modification.Applied microbiology and biotechnology · 2022Review
- Recombination of Compatible Substitutions by 2GenReP and InSiReP.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Multiparameter screening method for developing optimized red-fluorescent proteins.Nature protocols · 2020Article
- Computer-Assisted Recombination (CompassR) Teaches us How to Recombine Beneficial Substitutions from Directed Evolution Campaigns.Chemistry (Weinheim an der Bergstrasse, Germany) · 2020Article
- Golden Mutagenesis: An efficient multi-site-saturation mutagenesis approach by Golden Gate cloning with automated primer design.Scientific reports · 2019Article
- Development and Applications of Superfolder and Split Fluorescent Protein Detection Systems in Biology.International journal of molecular sciences · 2019Review
- Harnessing glycoenzyme engineering for synthesis of bioactive oligosaccharides.Interface focus · 2019Review
- Darwin Assembly: fast, efficient, multi-site bespoke mutagenesis.Nucleic acids research · 2018Article
- Article
- Modern methods for laboratory diversification of biomolecules.Current opinion in chemical biology · 2017Review
- mScarlet: a bright monomeric red fluorescent protein for cellular imaging.Nature methods · 2017Article
- In vitro flow cytometry-based screening platform for cellulase engineering.Scientific reports · 2016Article
- Heterologous expression of flax PHOSPHOLIPID:DIACYLGLYCEROL CHOLINEPHOSPHOTRANSFERASE (PDCT) increases polyunsaturated fatty acid content in yeast and Arabidopsis seeds.BMC biotechnology · 2015Article
- Combinatorial optimization of synthetic operons for the microbial production of p-coumaryl alcohol with Escherichia coli.Microbial cell factories · 2015Article
- Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently.Chemical Society reviews · 2015Review
- A high-throughput screening method to reengineer DNA polymerases for random mutagenesis.Molecular biotechnology · 2014Article
- Patch cloning method for multiple site-directed and saturation mutagenesis.BMC biotechnology · 2013Article
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4 authors.
Funding
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Abstract
Focused mutant library generation methods have been developed to improve mainly "localizable" enzyme properties such as activity and selectivity. Current multi-site saturation methods are restricted by the gene sequence, require subsequent PCR steps and/or additional enzymatic modifications. Here we report, a multiple site saturation mutagenesis method, OmniChange, which simultaneously and efficiently saturates five independent codons. As proof of principle, five chemically cleaved DNA fragments, each carrying one NNK-degenerated codon, were generated and assembled to full gene length in a one-pot-reaction without additional PCR-amplification or use of restriction enzymes or ligases. Sequencing revealed the presence of up to 27 different codons at individual positions, corresponding to 84.4% of the theoretical diversity offered by NNK-degeneration. OmniChange is absolutely sequence independent, does not require a minimal distance between mutated codons and can be accomplished within a day.
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