ArticleACS synthetic biology2017
The Impact of DNA Topology and Guide Length on Target Selection by a Cytosine-Specific Cas9.
Article in ACS synthetic biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed.
- Customizing CRISPR-Cas PAM specificity with protein language models.Nature biotechnology · 2026Article
- PAM-readID is a rapid, simple, and accurate PAM determination method for CRISPR-Cas enzymes in mammalian cells.Communications biology · 2025Article
- Insights into the compact CRISPR-Cas9d system.Nature communications · 2025Article
- Engineering of CRISPR-Cas PAM recognition using deep learning of vast evolutionary data.bioRxiv : the preprint server for biology · 2025Article
- Expression and Functional Analysis of the Compact ThermophilicInternational journal of molecular sciences · 2023Article
- A thermostable type I-B CRISPR-Cas system for orthogonal and multiplexed genetic engineering.Nature communications · 2023Article
- Coupled catalytic states and the role of metal coordination in Cas9.Nature catalysis · 2023Article
- The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.PLoS biology · 2021Article
- Efficient genome editing of an extreme thermophile, Thermus thermophilus, using a thermostable Cas9 variant.Scientific reports · 2021Article
- Discovery and engineering of small SlugCas9 with broad targeting range and high specificity and activity.Nucleic acids research · 2021Article
- The bridge helix of Cas12a imparts selectivity in cis-DNA cleavage and regulates trans-DNA cleavage.FEBS letters · 2021Article
- Catalytically Enhanced Cas9 through Directed Protein Evolution.The CRISPR journal · 2021Article
- CRISPR technologies and the search for the PAM-free nuclease.Nature communications · 2021Review
- Genome Editing of the Anaerobic Thermophile Thermoanaerobacter ethanolicus Using Thermostable Cas9.Applied and environmental microbiology · 2020Article
- The molecular basis for recognition of 5'-NNNCC-3' PAM and its methylation state by Acidothermus cellulolyticus Cas9.Nature communications · 2020Article
- Development of both type I-B and type II CRISPR/Cas genome editing systems in the cellulolytic bacteriumMetabolic engineering communications · 2020Article
- A compact Cas9 ortholog from Staphylococcus Auricularis (SauriCas9) expands the DNA targeting scope.PLoS biology · 2020Article
- Directed evolution studies of a thermophilic Type II-C Cas9.Methods in enzymology · 2019Article
- Phosphate Lock Residues of Acidothermus cellulolyticus Cas9 Are Critical to Its Substrate Specificity.ACS synthetic biology · 2018Article
- A CRISPR-Cas9-triggered strand displacement amplification method for ultrasensitive DNA detection.Nature communications · 2018Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Cas9 is an RNA-guided DNA cleavage enzyme being actively developed for genome editing and gene regulation. To be cleaved by Cas9, a double stranded DNA, or the protospacer, must be complementary to the guide region, typically 20-nucleotides in length, of the Cas9-bound guide RNA, and adjacent to a short Cas9-specific element called Protospacer Adjacent Motif (PAM). Understanding the correct juxtaposition of the protospacer- and PAM-interaction with Cas9 will enable development of versatile and safe Cas9-based technology. We report identification and biochemical characterization of Cas9 from Acidothermus cellulolyticus (AceCas9). AceCas9 depends on a 5'-NNNCC-3' PAM and is more efficient in cleaving negative supercoils than relaxed DNA. Kinetic as well as in vivo activity assays reveal that AceCas9 achieves optimal activity when combined with a guide RNA containing a 24-nucleotide complementarity region. The cytosine-specific, DNA topology-sensitive, and extended guide-dependent properties of AceCas9 may be explored for specific genome editing applications.
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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.