ArticleNucleic acids research2019
Allele-specific genome editing using CRISPR-Cas9 is associated with loss of heterozygosity in diploid yeast.
Article in Nucleic acids research, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- Progresses of transposon research inSynthetic and systems biotechnology · 2026Review
- Deconstruction ofbioRxiv : the preprint server for biology · 2026Article
- High-resolution genotype-free mapping of genetic variation with CRI-SPA-Map.Genome research · 2026Article
- Simplifying multiplex genome engineering in Saccharomyces cerevisiae with intron-mediated Random Assembly and INtegration (RAIN).FEMS yeast research · 2026Article
- High-resolution, genotype-free mapping of genetic variation with CRI-SPA-Map.bioRxiv : the preprint server for biology · 2025Article
- Homing gene drives can transfer rapidly between Anopheles gambiae strains with minimal carryover of flanking sequences.Nature communications · 2024Article
- Detection of unintended on-target effects in CRISPR genome editing by DNA donors carrying diagnostic substitutions.Nucleic acids research · 2023Article
- CRISPR-Cas9 engineering in the hybrid yeast Zygosaccharomyces parabailii can lead to loss of heterozygosity in target chromosomes.FEMS yeast research · 2023Article
- CRISPR-Cas9 Editing Induces Loss of Heterozygosity in the Pathogenic Yeast Candida parapsilosis.mSphere · 2022Article
- Differences in barriers for controlled learning about safety between biotechnology and chemistry.Nature communications · 2022Article
- Genomic Variation-Mediating Fluconazole Resistance in Yeast.Biomolecules · 2022Review
- Homozygous might be hemizygous: CRISPR/Cas9 editing in iPSCs results in detrimental on-target defects that escape standard quality controls.Stem cell reports · 2022Article
- A Saccharomyces eubayanus haploid resource for research studies.Scientific reports · 2022Article
- Tips, Tricks, and Potential Pitfalls of CRISPR Genome Editing inFrontiers in bioengineering and biotechnology · 2022Review
- Efficient breeding of industrial brewing yeast strains using CRISPR/Cas9-aided mating-type switching.Applied microbiology and biotechnology · 2021Article
- Responsible Learning About Risks Arising from Emerging Biotechnologies.Science and engineering ethics · 2021Article
- CRISPR-Cas9: A Powerful Tool to Efficiently EngineerLife (Basel, Switzerland) · 2020Review
- Mutation-Specific Guide RNA for Compound Heterozygous Porphyria On-target Scarless Correction by CRISPR/Cas9 in Stem Cells.Stem cell reports · 2020Article
- Plasmid-based complementation of large deletions in Phaeodactylum tricornutum biosynthetic genes generated by Cas9 editing.Scientific reports · 2020Article
- Safe-by-Design: Stakeholders' Perceptions and Expectations of How to Deal with Uncertain Risks of Emerging Biotechnologies in the Netherlands.Risk analysis : an official publication of the Society for Risk Analysis · 2020Article
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7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Targeted DNA double-strand breaks (DSBs) with CRISPR-Cas9 have revolutionized genetic modification by enabling efficient genome editing in a broad range of eukaryotic systems. Accurate gene editing is possible with near-perfect efficiency in haploid or (predominantly) homozygous genomes. However, genomes exhibiting polyploidy and/or high degrees of heterozygosity are less amenable to genetic modification. Here, we report an up to 99-fold lower gene editing efficiency when editing individual heterozygous loci in the yeast genome. Moreover, Cas9-mediated introduction of a DSB resulted in large scale loss of heterozygosity affecting DNA regions up to 360 kb and up to 1700 heterozygous nucleotides, due to replacement of sequences on the targeted chromosome by corresponding sequences from its non-targeted homolog. The observed patterns of loss of heterozygosity were consistent with homology directed repair. The extent and frequency of loss of heterozygosity represent a novel mutagenic side-effect of Cas9-mediated genome editing, which would have to be taken into account in eukaryotic gene editing. In addition to contributing to the limited genetic amenability of heterozygous yeasts, Cas9-mediated loss of heterozygosity could be particularly deleterious for human gene therapy, as loss of heterozygous functional copies of anti-proliferative and pro-apoptotic genes is a known path to cancer.
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