Matches in SemOpenAlex for { <https://semopenalex.org/work/W2954039297> ?p ?o ?g. }
- W2954039297 abstract "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." @default.
- W2954039297 created "2019-07-12" @default.
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- W2954039297 date "2019-07-04" @default.
- W2954039297 modified "2023-10-09" @default.
- W2954039297 title "Pooled clone collections by multiplexed CRISPR-Cas12a-assisted gene tagging in yeast" @default.
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- W2954039297 doi "https://doi.org/10.1038/s41467-019-10816-7" @default.
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