ArticleNature biotechnology2013
DNA targeting specificity of RNA-guided Cas9 nucleases.
Article in Nature biotechnology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2,742 papers, 2 of them syntheses that pooled it.
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Who cites it
2,742 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- The evolving landscape of gene editing therapies for human genetic diseases: a twenty-year bibliometric analysis.Frontiers in medicine · 2026Pooled it
- Antisense Versus Antigene in the Computer-Aided Design of Triplex-Forming Oligonucleotides (TFO): Insights from a Dual-Method Review, Combining Bibliometric and Systematic Review.International journal of molecular sciences · 2025Pooled it
- Adeno-Associated Viral Vector (AAV)-Mediated In Vivo CRISPR-Cas9 Delivery.Methods in molecular biology (Clifton, N.J.) · 2027Article
- Digenome-Detect: Accurate, statistics-based analysis software for identifying off-target cleavage sites in genome editing.Molecular therapy. Advances · 2026Article
- Transposon insertion sequencing identifies novel genes involved in product synthesis in gas-fermentingSynthetic and systems biotechnology · 2026Article
- Translating base editors: a decade of progress from the bench to the clinic.Nature reviews. Drug discovery · 2026Review
- Modeling patient variants of Cnot1 and Cdc42bpb results in distinct forms of congenital diaphragmatic hernia in mice.Human molecular genetics · 2026Article
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- BSA-seq and fine linkage mapping for the identification of a novel locus (qNUE6) for nitrogen use efficiency in rice.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2026Article
- Minimizing Off-Target Effects of CRISPR-Cas9 With Optimized sgRNA: Evaluation of Efficiency and Specificity in the Tumor Protein 53 (TP53) Region.Biotechnology and bioengineering · 2026Article
- An iron-regulated methionine redox axis governs adipose browning and cancer cachexia.Nature cancer · 2026Article
- Generation ofJBMR plus · 2026Article
- Rational selection of SpCas9 and variant-derived adenine base editors for human genome editing.Gene therapy · 2026Article
- RNA therapeutics: current status and future directions.Signal transduction and targeted therapy · 2026Review
- Auto-SMART: a web-based platform for repair-template design for CRISPR/Cas9 homology-directed editing.bioRxiv : the preprint server for biology · 2026Article
- A dynamically frustrated allosteric checkpoint consistent with conformational proofreading in CRISPR-Cas9.Nucleic acids research · 2026Article
- Size and sequence variation of the Lid of the Cas12a nuclease domain results in a nickase phenotype.Nucleic acids research · 2026Article
- A high-throughput co-culture CRISPR array for rapid fitness hypothesis testing and growth engineering in CHO cells.iScience · 2026Article
- Active-site arginines differentially control Cas12a DNA cleavage and specificity.The Journal of biological chemistry · 2026Article
2,682 more citing papers are in PubMed but not listed here.
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14 authors.
Funding
Abstract
The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by means of single-guide RNAs (sgRNAs) to enable genome editing. Here, we characterize SpCas9 targeting specificity in human cells to inform the selection of target sites and avoid off-target effects. Our study evaluates >700 guide RNA variants and SpCas9-induced indel mutation levels at >100 predicted genomic off-target loci in 293T and 293FT cells. We find that SpCas9 tolerates mismatches between guide RNA and target DNA at different positions in a sequence-dependent manner, sensitive to the number, position and distribution of mismatches. We also show that SpCas9-mediated cleavage is unaffected by DNA methylation and that the dosage of SpCas9 and sgRNA can be titrated to minimize off-target modification. To facilitate mammalian genome engineering applications, we provide a web-based software tool to guide the selection and validation of target sequences as well as off-target analyses.
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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.