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Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B

To fend off foreign genetic elements, prokaryotes have developed several defense systems. The most recently discovered defense system, CRISPR/Cas, is sequence-specific, adaptive and heritable. The two central components of this system are the Cas proteins and the CRISPR RNA. The latter consists of r...

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Autores principales: Maier, Lisa-Katharina, Lange, Sita J., Stoll, Britta, Haas, Karina A., Fischer, Susan, Fischer, Eike, Duchardt-Ferner, Elke, Wöhnert, Jens, Backofen, Rolf, Marchfelder, Anita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Landes Bioscience 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737343/
https://www.ncbi.nlm.nih.gov/pubmed/23594992
http://dx.doi.org/10.4161/rna.24282
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author Maier, Lisa-Katharina
Lange, Sita J.
Stoll, Britta
Haas, Karina A.
Fischer, Susan
Fischer, Eike
Duchardt-Ferner, Elke
Wöhnert, Jens
Backofen, Rolf
Marchfelder, Anita
author_facet Maier, Lisa-Katharina
Lange, Sita J.
Stoll, Britta
Haas, Karina A.
Fischer, Susan
Fischer, Eike
Duchardt-Ferner, Elke
Wöhnert, Jens
Backofen, Rolf
Marchfelder, Anita
author_sort Maier, Lisa-Katharina
collection PubMed
description To fend off foreign genetic elements, prokaryotes have developed several defense systems. The most recently discovered defense system, CRISPR/Cas, is sequence-specific, adaptive and heritable. The two central components of this system are the Cas proteins and the CRISPR RNA. The latter consists of repeat sequences that are interspersed with spacer sequences. The CRISPR locus is transcribed into a precursor RNA that is subsequently processed into short crRNAs. CRISPR/Cas systems have been identified in bacteria and archaea, and data show that many variations of this system exist. We analyzed the requirements for a successful defense reaction in the halophilic archaeon Haloferax volcanii. Haloferax encodes a CRISPR/Cas system of the I-B subtype, about which very little is known. Analysis of the mature crRNAs revealed that they contain a spacer as their central element, which is preceded by an eight-nucleotide-long 5′ handle that originates from the upstream repeat. The repeat sequences have the potential to fold into a minimal stem loop. Sequencing of the crRNA population indicated that not all of the spacers that are encoded by the three CRISPR loci are present in the same abundance. By challenging Haloferax with an invader plasmid, we demonstrated that the interaction of the crRNA with the invader DNA requires a 10-nucleotide-long seed sequence. In addition, we found that not all of the crRNAs from the three CRISPR loci are effective at triggering the degradation of invader plasmids. The interference does not seem to be influenced by the copy number of the invader plasmid.
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spelling pubmed-37373432013-08-28 Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B Maier, Lisa-Katharina Lange, Sita J. Stoll, Britta Haas, Karina A. Fischer, Susan Fischer, Eike Duchardt-Ferner, Elke Wöhnert, Jens Backofen, Rolf Marchfelder, Anita RNA Biol Research Paper To fend off foreign genetic elements, prokaryotes have developed several defense systems. The most recently discovered defense system, CRISPR/Cas, is sequence-specific, adaptive and heritable. The two central components of this system are the Cas proteins and the CRISPR RNA. The latter consists of repeat sequences that are interspersed with spacer sequences. The CRISPR locus is transcribed into a precursor RNA that is subsequently processed into short crRNAs. CRISPR/Cas systems have been identified in bacteria and archaea, and data show that many variations of this system exist. We analyzed the requirements for a successful defense reaction in the halophilic archaeon Haloferax volcanii. Haloferax encodes a CRISPR/Cas system of the I-B subtype, about which very little is known. Analysis of the mature crRNAs revealed that they contain a spacer as their central element, which is preceded by an eight-nucleotide-long 5′ handle that originates from the upstream repeat. The repeat sequences have the potential to fold into a minimal stem loop. Sequencing of the crRNA population indicated that not all of the spacers that are encoded by the three CRISPR loci are present in the same abundance. By challenging Haloferax with an invader plasmid, we demonstrated that the interaction of the crRNA with the invader DNA requires a 10-nucleotide-long seed sequence. In addition, we found that not all of the crRNAs from the three CRISPR loci are effective at triggering the degradation of invader plasmids. The interference does not seem to be influenced by the copy number of the invader plasmid. Landes Bioscience 2013-05-01 2013-04-17 /pmc/articles/PMC3737343/ /pubmed/23594992 http://dx.doi.org/10.4161/rna.24282 Text en Copyright © 2013 Landes Bioscience http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.
spellingShingle Research Paper
Maier, Lisa-Katharina
Lange, Sita J.
Stoll, Britta
Haas, Karina A.
Fischer, Susan
Fischer, Eike
Duchardt-Ferner, Elke
Wöhnert, Jens
Backofen, Rolf
Marchfelder, Anita
Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title_full Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title_fullStr Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title_full_unstemmed Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title_short Essential requirements for the detection and degradation of invaders by the Haloferax volcanii CRISPR/Cas system I-B
title_sort essential requirements for the detection and degradation of invaders by the haloferax volcanii crispr/cas system i-b
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3737343/
https://www.ncbi.nlm.nih.gov/pubmed/23594992
http://dx.doi.org/10.4161/rna.24282
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