Abstract
We developed a CRISPR-Cas9- and homology-directed-repair-assisted genome-scale engineering method named CHAnGE that can rapidly output tens of thousands of specific genetic variants in yeast. More than 98% of target sequences were efficiently edited with an average frequency of 82%. We validate the single-nucleotide resolution genome-editing capability of this technology by creating a genome-wide gene disruption collection and apply our method to improve tolerance to growth inhibitors.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Validation Study
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Biotechnology
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CRISPR-Cas Systems
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DNA, Fungal / genetics
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Directed Molecular Evolution
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Gene Editing / methods
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Genetic Engineering / methods*
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Genome, Fungal
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Models, Molecular
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Mutagenesis
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Recombinational DNA Repair
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Ubiquitin-Protein Ligases / chemistry
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Ubiquitin-Protein Ligases / genetics
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Ubiquitin-Protein Ligases / metabolism
Substances
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DNA, Fungal
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Saccharomyces cerevisiae Proteins
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Ubiquitin-Protein Ligases
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Siz1 protein, S cerevisiae