ArticleCurrent genetics2014
Rapid, efficient and precise allele replacement in the fission yeast Schizosaccharomyces pombe.
Article in Current genetics, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed.
- Proteasome regulation of petite-negativity in fission yeast.BMC biology · 2025Article
- Automated plasmid design for marker-free genome editing in budding yeast.G3 (Bethesda, Md.) · 2025Article
- The nuclear poly(A)-binding protein Pab2/PABPN1 promotes heterochromatin assembly through the formation of Pab2 nuclear condensates.PLoS genetics · 2025Article
- Targeted Forward Genetics: Saturating Mutational Analyses of Specific Target Loci Within the Genome.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Long multiply marked DNA repair template reveals lengths and fidelity of genome editing tracts inmicroPublication biology · 2025Article
- Investigating Fission Yeast Mutagenesis Using Canavanine Sensitivity Assays.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Agar lot-specific inhibition in the plating efficiency of yeast spores and cells.G3 (Bethesda, Md.) · 2024Article
- Creating Meiotic Recombination-Regulating DNA Sites byBiomolecules · 2024Article
- Laboratory horror stories: Poison in the agars.bioRxiv : the preprint server for biology · 2024Article
- Distance-dependent effects on CRISPR/Cas9-mediated genome editing inmicroPublication biology · 2024Article
- A marker-free genome editing method inmicroPublication biology · 2023Article
- Identification of an integrase-independent pathway of retrotransposition.Science advances · 2022Article
- Targeted Forward Genetics: Population-Scale Analyses of Allele Replacements Spanning Thousands of Base Pairs in Fission Yeast.G3 (Bethesda, Md.) · 2019Article
- Heterochromatin suppresses gross chromosomal rearrangements at centromeres by repressing Tfs1/TFIIS-dependent transcription.Communications biology · 2019Article
- Structure of the replication regulator Sap1 reveals functionally important interfaces.Scientific reports · 2018Article
- Histone deacetylation promotes transcriptional silencing at facultative heterochromatin.Nucleic acids research · 2018Article
- Tight Regulation of Srs2 Helicase Activity Is Crucial for Proper Functioning of DNA Repair Mechanisms.G3 (Bethesda, Md.) · 2018Article
- TORC1 and TORC2 converge to regulate the SAGA co-activator in response to nutrient availability.EMBO reports · 2017Article
- Metabolic engineering ofMetabolic engineering communications · 2017Article
- Big data mining powers fungal research: recent advances in fission yeast systems biology approaches.Current genetics · 2017Review
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Gene targeting provides a powerful tool to modify endogenous loci to contain specific mutations, insertions and deletions. Precise allele replacement, with no other chromosomal changes (e.g., insertion of selectable markers or heterologous promoters), maintains physiologically relevant context. Established methods for precise allele replacement in fission yeast employ two successive rounds of transformation and homologous recombination and require genotyping at each step. The relative efficiency of homologous recombination is low and a high rate of false positives during the second round of gene targeting further complicates matters. We report that pop-in, pop-out allele replacement circumvents these problems. We present data for 39 different allele replacements, involving simple and complex modifications at seven different target loci, that illustrate the power and utility of the approach. We also developed and validated a rapid, efficient process for precise allele replacement that requires only one round each of transformation and genotyping. We show that this process can be applied in population scale to an individual target locus, without genotyping, to identify clones with an altered phenotype (targeted forward genetics). It is therefore suitable for saturating, in situ, locus-specific mutation screens (e.g., of essential or non-essential genes and regulatory DNA elements) within normal chromosomal context.
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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.