ArticleNature communications2019
Pooled clone collections by multiplexed CRISPR-Cas12a-assisted gene tagging in yeast.
Article in Nature communications, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Brazilian Biomes as Promising Resources of Rhodotorula Yeasts for the Biotechnological Production of Carotenoids.Chemistry & biodiversity · 2025Review
- High throughput mutagenesis and screening for yeast engineering.Journal of biological engineering · 2022Review
- Characterizing Endogenous Protein Complexes with Biological Mass Spectrometry.Chemical reviews · 2022Review
- Applications of CRISPR/Cas gene-editing technology in yeast and fungi.Archives of microbiology · 2021Article
- Analysis of local protein accumulation kinetics by live-cell imaging in yeast systems.STAR protocols · 2021Article
- A colorimetric RT-LAMP assay and LAMP-sequencing for detecting SARS-CoV-2 RNA in clinical samples.Science translational medicine · 2020Article
- CRISPR-Cas12a-assisted PCR tagging of mammalian genes.The Journal of cell biology · 2020Article
- Article
- Design and Construction of Portable CRISPR-Cpf1-Mediated Genome Editing inFrontiers in bioengineering and biotechnology · 2020Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Clone collections of modified strains ("libraries") are a major resource for systematic studies with the yeast Saccharomyces cerevisiae. Construction of such libraries is time-consuming, costly and confined to the genetic background of a specific yeast strain. To overcome these limitations, we present CRISPR-Cas12a (Cpf1)-assisted tag library engineering (CASTLING) for multiplexed strain construction. CASTLING uses microarray-synthesized oligonucleotide pools and in vitro recombineering to program the genomic insertion of long DNA constructs via homologous recombination. One simple transformation yields pooled libraries with >90% of correctly tagged clones. Up to several hundred genes can be tagged in a single step and, on a genomic scale, approximately half of all genes are tagged with only ~10-fold oversampling. We report several parameters that affect tagging success and provide a quantitative targeted next-generation sequencing method to analyze such pooled collections. Thus, CASTLING unlocks avenues for increasing throughput in functional genomics and cell biology research.
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Registered trials
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