ArticleNucleic acids research2019
CRISPR repeat sequences and relative spacing specify DNA integration by Pyrococcus furiosus Cas1 and Cas2.
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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Who cites it
20 citing papers in PubMed, 23 citations in OpenAlex.
- Defense Systems and Prophage Detection in Streptococcus mutans Strains.Molecular oral microbiology · 2026Article
- Insights into spacer acquisition of the type V-A CRISPR-Cas system of Francisella novicida U112.Nucleic acids research · 2026Article
- CRISPR-Cas Systems: A Functional Perspective and Innovations.International journal of molecular sciences · 2025Review
- Histones direct site-specific CRISPR spacer acquisition in model archaeon.Nature microbiology · 2023Article
- DNA Motifs and an Accessory CRISPR Factor Determine Cas1 Binding and Integration Activity inInternational journal of molecular sciences · 2022Article
- Unique properties of spacer acquisition by the type III-A CRISPR-Cas system.Nucleic acids research · 2022Article
- Transcriptional analysis of CRISPR I-B arrays ofFrontiers in microbiology · 2022Article
- Adaptation by Type III CRISPR-Cas Systems: Breakthrough Findings and Open Questions.Frontiers in microbiology · 2022Review
- CRISPR Gene Editing in Lipid Disorders and Atherosclerosis: Mechanisms and Opportunities.Metabolites · 2021Review
- Sequence specific integration by the family 1 casposase from Candidatus Nitrosopumilus koreensis AR1.Nucleic acids research · 2021Article
- Virus-induced cell gigantism and asymmetric cell division in archaea.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Novel Strategy to Combat Antibiotic Resistance: A Sight into the Combination of CRISPR/Cas9 and Nanoparticles.Pharmaceutics · 2021Review
- Pruning and Tending Immune Memories: Spacer Dynamics in the CRISPR Array.Frontiers in microbiology · 2021Review
- Heavily Armed Ancestors: CRISPR Immunity and Applications in Archaea with a Comparative Analysis of CRISPR Types in Sulfolobales.Biomolecules · 2020Review
- Primed CRISPR DNA uptake in Pyrococcus furiosus.Nucleic acids research · 2020Article
- Atypical organizations and epistatic interactions of CRISPRs and cas clusters in genomes and their mobile genetic elements.Nucleic acids research · 2020Article
- Growth inhibition by bacterial Cas2Em proteins expressed in mammalian cells.American journal of translational research · 2020Article
- Fidelity of prespacer capture and processing is governed by the PAM-mediated interactions of Cas1-2 adaptation complex in CRISPR-Cas type I-E system.The Journal of biological chemistry · 2019Article
- CRISPR DNA elements controlling site-specific spacer integration and proper repeat length by a Type II CRISPR-Cas system.Nucleic acids research · 2019Article
- The Application of the CRISPR/Cas9 System in the Treatment of Hepatitis B Liver Cancer.Technology in cancer research & treatmentReview
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
Acquiring foreign spacer DNA into the CRISPR locus is an essential primary step of the CRISPR-Cas pathway in prokaryotes for developing host immunity to mobile genetic elements. Here, we investigate spacer integration in vitro using proteins from Pyrococcus furiosus and demonstrate that Cas1 and Cas2 are sufficient to accurately integrate spacers into a minimal CRISPR locus. Using high-throughput sequencing, we identified high frequency spacer integration occurring at the same CRISPR repeat border sites utilized in vivo, as well as at several non-CRISPR plasmid sequences which share features with repeats. Analysis of non-CRISPR integration sites revealed that Cas1 and Cas2 are directed to catalyze full-site spacer integration at specific DNA stretches where guanines and/or cytosines are 30 base pairs apart and the intervening sequence harbors several positionally conserved bases. Moreover, assaying a series of CRISPR repeat mutations, followed by sequencing of the integration products, revealed that the specificity of integration is primarily directed by sequences at the leader-repeat junction as well as an adenine-rich sequence block in the mid-repeat. Together, our results indicate that P. furiosus Cas1 and Cas2 recognize multiple sequence features distributed over a 30 base pair DNA region for accurate spacer integration at the CRISPR repeat.
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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.