ArticleYeast (Chichester, England)2014
The 50:50 method for PCR-based seamless genome editing in yeast.
Article in Yeast (Chichester, England), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- PYEAST - A Computational Toolkit for Saccharomyces cerevisiae Genetic Engineering.NPJ systems biology and applications · 2026Article
- Automated plasmid design for marker-free genome editing in budding yeast.G3 (Bethesda, Md.) · 2025Article
- Free introns of tRNAs as complementarity-dependent regulators of gene expression.Molecular cell · 2025Article
- Pooled PPIseq: Screening the SARS-CoV-2 and human interface with a scalable multiplexed protein-protein interaction assay platform.PloS one · 2025Article
- Cytoplasmic redox imbalance in the thioredoxin system activates Hsf1 and results in hyperaccumulation of the sequestrase Hsp42 with misfolded proteins.Molecular biology of the cell · 2024Article
- Up-regulation of ubiquitin-proteasome activity upon loss of NatA-dependent N-terminal acetylation.Life science alliance · 2022Article
- Article
- Wheat Encodes Small, Secreted Proteins That Contribute to Resistance to Septoria Tritici Blotch.Frontiers in genetics · 2020Article
- Rap1-mediated nucleosome displacement can regulate gene expression in senescent cells without impacting the pace of senescence.Aging cell · 2020Article
- Molecular interactions between Hel2 and RNA supporting ribosome-associated quality control.Nature communications · 2019Article
- Engineered Biosensors from Dimeric Ligand-Binding Domains.ACS synthetic biology · 2018Article
- HEx: A heterologous expression platform for the discovery of fungal natural products.Science advances · 2018Article
- A method for high-throughput production of sequence-verified DNA libraries and strain collections.Molecular systems biology · 2017Article
- Ndt80 activates the meiotic ORC1 transcript isoform and SMA2 via a bi-directional middle sporulation element in Saccharomyces cerevisiae.RNA biology · 2016Article
- Article
- Yeast as a cell factory: current state and perspectives.Microbial cell factories · 2015Review
- Proteasomes, Sir2, and Hxk2 form an interconnected aging network that impinges on the AMPK/Snf1-regulated transcriptional repressor Mig1.PLoS genetics · 2015Article
- The conserved histone deacetylase Rpd3 and its DNA binding subunit Ume6 control dynamic transcript architecture during mitotic growth and meiotic development.Nucleic acids research · 2015Article
- Scalable and versatile genome editing using linear DNAs with microhomology to Cas9 Sites in Caenorhabditis elegans.Genetics · 2014Article
- IpO: plasmids and methods for simplified, PCR-based DNA transplant in yeast.Yeast (Chichester, England) · 2014Article
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2 authors.
Funding
Abstract
The ability to edit the yeast genome with relative ease has contributed to the organism being a model eukaryote for decades. Most methods for deleting, inserting or altering genomic sequences require transformation with DNA that carries the desired change and a selectable marker. One-step genome editing methods retain the selectable marker. Seamless genome editing methods require more steps and a marker that can be used for both positive and negative selection, such as URA3. Here we describe the PCR-based 50:50 method for seamless genome editing, which requires only two primers, one PCR with a URA3 cassette, and a single yeast transformation. Our method is based on pop-in/pop-out gene replacement and is amenable to the facile creation of genomic deletions and short insertions or substitutions. We used the 50:50 method to make two conservative loss-of-function mutations in MATα1, with results suggesting that the wild-type gene has a new function outside of that presently known.
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