ArticlePloS one2017
A method for labeling proteins with tags at the native genomic loci in budding yeast.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Zinc-Finger 5 Is an Activation Domain in theJournal of fungi (Basel, Switzerland) · 2025Article
- Exploring CRISPR/Cas9-Mediated Gene Editing Advances in Conventional and Non-conventional Yeast Species.Applied biochemistry and biotechnology · 2025Review
- Article
- Transcriptional activation of budding yeast DDI2/3 through chemical modifications of Fzf1.Cell biology and toxicology · 2023Article
- An adaptable live-cell imaging protocol to analyze organelle morphology inSTAR protocols · 2022Article
- Charged Polypeptide Tail Boosts the Salt Resistance of Enzyme-Containing Complex Coacervate Micelles.Biomacromolecules · 2022Article
- Cell Synchronization Enhances Nuclear Transformation and Genome EditingACS synthetic biology · 2020Article
- Fine-tuning the expression of target genes using a DDI2 promoter gene switch in budding yeast.Scientific reports · 2019Article
- Marker-free genetic manipulations in yeast using CRISPR/CAS9 system.Current genetics · 2018Article
- CRISPR-UnLOCK: Multipurpose Cas9-Based Strategies for Conversion of Yeast Libraries and Strains.Frontiers in microbiology · 2017Article
- Utilizing Biotinylated Proteins Expressed in Yeast to Visualize DNA-Protein Interactions at the Single-Molecule Level.Frontiers in microbiology · 2017Article
Corrections and comments
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique in the budding yeast Saccharomyces cerevisiae is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. However, several limitations hamper the application of this technique, such as the selectable markers not being reusable, tagging of only the C-terminal being possible, and a "scar" sequence being left in the genome. Here, we describe a strategy to solve these problems by tagging target genes based on a pop-in/pop-out and counter-selection system. Three fluorescent protein tag (mCherry, sfGFP, and mKikGR) and two epitope tag (HA and 3×FLAG) constructs were developed and utilized to tag HHT1, UBC13 or RAD5 at the chromosomal locus as proof-of-concept.
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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.