ArticleNucleic acids research2017
Structural constraints and enzymatic promiscuity in the Cas6-dependent generation of crRNAs.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Deletion of the gene for a cyanobacterial ribosome-associated protein affects the carbon/nitrogen metabolism.bioRxiv : the preprint server for biology · 2026Article
- Comprehensive analysis of CRISPR array repeat mutations reveals subtype-specific patterns and links to spacer dynamics.microLife · 2026Article
- The CRISPR-cas repertoire of Kluyvera ascorbata: insights from genomic data.Current genetics · 2025Article
- Article
- Cyanobacterial type I CRISPR-Cas systems: distribution, mechanisms, and genome editing applications.Frontiers in bioengineering and biotechnology · 2025Review
- RNA targeting and cleavage by the type III-Dv CRISPR effector complex.Nature communications · 2024Article
- Shortened CRISPR-Cas9 arrays enable multiplexed gene targeting in bacteria from a smaller DNA footprint.RNA biology · 2023Article
- Regulation of pSYSA defense plasmid copy number inFrontiers in microbiology · 2023Article
- Enzymatic properties of CARF-domain proteins inFrontiers in microbiology · 2022Article
- Transcriptional analysis of CRISPR I-B arrays ofFrontiers in microbiology · 2022Article
- The tracrRNA in CRISPR Biology and Technologies.Annual review of genetics · 2021Review
- CRISPRloci: comprehensive and accurate annotation of CRISPR-Cas systems.Nucleic acids research · 2021Article
- Analysis of a photosynthetic cyanobacterium rich in internal membrane systems via gradient profiling by sequencing (Grad-seq).The Plant cell · 2021Article
- CRISPRidentify: identification of CRISPR arrays using machine learning approach.Nucleic acids research · 2021Article
- Specificities and functional coordination between the two Cas6 maturation endonucleases inRNA biology · 2020Article
- Divergent methylation of CRISPR repeats and cas genes in a subtype I-D CRISPR-Cas-system.BMC microbiology · 2019Article
- Biochemical analysis of the Cas6-1 RNA endonuclease associated with the subtype I-D CRISPR-Cas system in Synechocystis sp. PCC 6803.RNA biology · 2019Article
- The nuts and bolts of the Haloferax CRISPR-Cas system I-B.RNA biology · 2019Article
- Comprehensive search for accessory proteins encoded with archaeal and bacterial type III CRISPR-cas gene cassettes reveals 39 new cas gene families.RNA biology · 2019Article
- CRISPR-Cas systems in multicellular cyanobacteria.RNA biology · 2019Article
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7 authors.
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No grant is acknowledged in the PubMed record.
Abstract
A hallmark of defense mechanisms based on clustered regularly interspaced short palindromic repeats (CRISPR) and associated sequences (Cas) are the crRNAs that guide these complexes in the destruction of invading DNA or RNA. Three separate CRISPR-Cas systems exist in the cyanobacterium Synechocystis sp. PCC 6803. Based on genetic and transcriptomic evidence, two associated endoribonucleases, Cas6-1 and Cas6-2a, were postulated to be involved in crRNA maturation from CRISPR1 or CRISPR2, respectively. Here, we report a promiscuity of both enzymes to process in vitro not only their cognate transcripts, but also the respective non-cognate precursors, whereas they are specific in vivo Moreover, while most of the repeats serving as substrates were cleaved in vitro, some were not. RNA structure predictions suggested that the context sequence surrounding a repeat can interfere with its stable folding. Indeed, structure accuracy calculations of the hairpin motifs within the repeat sequences explained the majority of analyzed cleavage reactions, making this a good measure for predicting successful cleavage events. We conclude that the cleavage of CRISPR1 and CRISPR2 repeat instances requires a stable formation of the characteristic hairpin motif, which is similar between the two types of repeats. The influence of surrounding sequences might partially explain variations in crRNA abundances and should be considered when designing artificial CRISPR arrays.
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