ArticleMicrobial cell factories2015
CrEdit: CRISPR mediated multi-loci gene integration in Saccharomyces cerevisiae.
Article in Microbial cell factories, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 68 papers.
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Who cites it
68 citing papers in PubMed, 154 citations in OpenAlex.
- Parallel gene amplification by Cas9 nickase for generating functionally heterogeneous cell populations.Cell reports methods · 2026Article
- Exploring chromosomal position effects for predictable tuning of metabolic pathways in yeast.Nucleic acids research · 2026Article
- A toolkit for facilitating markerless integration of expression cassettes in Komagataella phaffii via CRISPR/Cas9.Microbial cell factories · 2025Article
- Strategic targeting of Cas9 nickase expands tandem gene arrays.Cell genomics · 2025Article
- Strategic targeting of Cas9 nickase induces large segmental duplications.Cell genomics · 2024Article
- Advances in yeast synthetic biology for human G protein-coupled receptor biology and pharmacology.Current opinion in biotechnology · 2024Review
- Multiplex CRISPR-Cas Genome Editing: Next-Generation Microbial Strain Engineering.Journal of agricultural and food chemistry · 2024Review
- The expanded CRISPR toolbox for constructing microbial cell factories.Trends in biotechnology · 2024Review
- A Multiplex MoClo Toolkit for Extensive and Flexible Engineering ofACS synthetic biology · 2023Article
- Practical Approaches for the YeastInternational journal of molecular sciences · 2023Review
- Acceleration of CRISPR/Cas9-Mediated Editing at Multiple Sites in theMethods and protocols · 2023Article
- Optimizing the strain engineering process for industrial-scale production of bio-based molecules.Journal of industrial microbiology & biotechnology · 2023Review
- pCEC-red: a new vector for easier and faster CRISPR-Cas9 genome editing in Saccharomyces cerevisiae.FEMS yeast research · 2023Article
- ACtivE: Assembly and CRISPR-TargetedACS synthetic biology · 2022Article
- Standardization of Synthetic Biology Tools and Assembly Methods forACS synthetic biology · 2022Review
- How adaptive laboratory evolution can boost yeast tolerance to lignocellulosic hydrolyses.Current genetics · 2022Review
- The MyLO CRISPR-Cas9 toolkit: a markerless yeast localization and overexpression CRISPR-Cas9 toolkit.G3 (Bethesda, Md.) · 2022Article
- Easy efficient HDR-based targeted knock-in inBioengineered · 2022Article
- Tips, Tricks, and Potential Pitfalls of CRISPR Genome Editing inFrontiers in bioengineering and biotechnology · 2022Review
- Applications of CRISPR/Cas gene-editing technology in yeast and fungi.Archives of microbiology · 2021Article
8 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
10 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundOne of the bottlenecks in production of biochemicals and pharmaceuticals in Saccharomyces cerevisiae is stable and homogeneous expression of pathway genes. Integration of genes into the genome of the production organism is often a preferred option when compared to expression from episomal vectors. Existing approaches for achieving stable simultaneous genome integrations of multiple DNA fragments often result in relatively low integration efficiencies and furthermore rely on the use of selection markers.
resultsHere, we have developed a novel method, CrEdit (CRISPR/Cas9 mediated genome Editing), which utilizes targeted double strand breaks caused by CRISPR/Cas9 to significantly increase the efficiency of homologous integration in order to edit and manipulate genomic DNA. Using CrEdit, the efficiency and locus specificity of targeted genome integrations reach close to 100% for single gene integration using short homology arms down to 60 base pairs both with and without selection. This enables direct and cost efficient inclusion of homology arms in PCR primers. As a proof of concept, a non-native β-carotene pathway was reconstructed in S. cerevisiae by simultaneous integration of three pathway genes into individual intergenic genomic sites. Using longer homology arms, we demonstrate highly efficient and locus-specific genome integration even without selection with up to 84% correct clones for simultaneous integration of three gene expression cassettes.
conclusionsThe CrEdit approach enables fast and cost effective genome integration for engineering of S. cerevisiae. Since the choice of the targeting sites is flexible, CrEdit is a powerful tool for diverse genome engineering applications.
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