ArticleNucleic acids research2019
R-loop formation by dCas9 is mutagenic in Saccharomyces cerevisiae.
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 20 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
20 citing papers in PubMed, 42 citations in OpenAlex.
- When R-Loops Go Awry: Genome Instability and Neurological Diseases.The European journal of neuroscience · 2026Review
- Live genome imaging by CRISPR engineering: progress and problems.Experimental & molecular medicine · 2025Review
- R-loops acted on by RNase H1 influence DNA replication timing and genome stability in Leishmania.Nature communications · 2025Article
- Modern Plant Breeding Techniques in Crop Improvement and Genetic Diversity: From Molecular Markers and Gene Editing to Artificial Intelligence-A Critical Review.Plants (Basel, Switzerland) · 2024Review
- A dynamic subpopulation of CRISPR-Cas overexpressers allows Streptococcus pyogenes to rapidly respond to phage.Nature microbiology · 2024Article
- Induced proximity labeling and editing for epigenetic research.Cell chemical biology · 2024Review
- CRISPR/Cas9 as a Mutagenic Factor.International journal of molecular sciences · 2024Review
- A Rapid Antibody Enhancement Platform inACS synthetic biology · 2023Article
- Toward the Development of Epigenome Editing-Based Therapeutics: Potentials and Challenges.International journal of molecular sciences · 2023Review
- DNA nicks induce mutational signatures associated with BRCA1 deficiency.Nature communications · 2022Article
- Systematic decomposition of sequence determinants governing CRISPR/Cas9 specificity.Nature communications · 2022Article
- Article
- Tips, Tricks, and Potential Pitfalls of CRISPR Genome Editing inFrontiers in bioengineering and biotechnology · 2022Review
- Targeted epigenetic repression by CRISPR/dSaCas9 suppresses pathogenicMolecular therapy. Methods & clinical development · 2021Article
- Catalytically inactive Cas9 impairs DNA replication fork progression to induce focal genomic instability.Nucleic acids research · 2021Article
- Replisome bypass of a protein-based R-loop block by Pif1.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Atypical UV Photoproducts Induce Non-canonical Mutation Classes Associated with Driver Mutations in Melanoma.Cell reports · 2020Article
- Zooming in on chromosome dynamics.Cell cycle (Georgetown, Tex.) · 2020Review
- Review
- Point mutations in topoisomerase I alter the mutation spectrum in E. coli and impact the emergence of drug resistance genotypes.Nucleic acids research · 2020Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Cas9 binds and cleaves specific DNA sequences by inducing the formation of an R-loop between the guide RNA and its genomic target. While targeting of active Cas9 to a genomic locus is highly mutagenic because Cas9 creates DNA double strand breaks, targeting of dead Cas9 (dCas9) is presumed not to be mutagenic, as dCas9 lacks DNA endonuclease activity. Here, we show that dCas9 targeting induces mutations in yeast, particularly when targeted to the non-transcribed strand of a gene. dCas9-induced mutations cluster near the guide RNA target region and are comprised of single nucleotide substitutions, small insertions and deletions, and even complex mutations, depending upon the particular guide RNA target. We show that many of these mutations are a consequence of cytosine deamination events occurring on the non-target strand of the dCas9-induced R-loop, while others are associated with homopolymer instability or translesion DNA synthesis. Targeting of dCas9 by a mismatch-containing guide RNA also increases CAN1 mutation frequency, particularly in an ung1Δ mutant strain, suggesting that dCas9 induces mutations through similar mechanisms at off-target sites. These findings indicate that DNA binding by dCas9 is mutagenic in yeast, likely because dCas9 induces the formation of an R-loop at its target site.
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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.