Cargando…

A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease

BACKGROUND: Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR. METHODS: The CRI...

Descripción completa

Detalles Bibliográficos
Autores principales: Ivanov, Yury V., Shariat, Nikki, Register, Karen B., Linz, Bodo, Rivera, Israel, Hu, Kai, Dudley, Edward G., Harvill, Eric T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4624362/
https://www.ncbi.nlm.nih.gov/pubmed/26502932
http://dx.doi.org/10.1186/s12864-015-2028-9
_version_ 1782397802697981952
author Ivanov, Yury V.
Shariat, Nikki
Register, Karen B.
Linz, Bodo
Rivera, Israel
Hu, Kai
Dudley, Edward G.
Harvill, Eric T.
author_facet Ivanov, Yury V.
Shariat, Nikki
Register, Karen B.
Linz, Bodo
Rivera, Israel
Hu, Kai
Dudley, Edward G.
Harvill, Eric T.
author_sort Ivanov, Yury V.
collection PubMed
description BACKGROUND: Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR. METHODS: The CRISPR-Cas system has been identified using Basic Local Alignment Search Tool (BLAST) against other sequenced and annotated genomes and confirmed via CRISPRfinder program. Using Polymerase Chain Reactions (PCR) and Sanger DNA sequencing, we discovered CRISPRs in additional bacterial isolates of the same species of Bordetella. Transcriptional activity and processing of the CRISPR have been assessed via RT-PCR. RESULTS: Here we describe a novel Type II-C CRISPR and its associated genes—cas1, cas2, and cas9—in several isolates of a newly discovered Bordetella species. The CRISPR-cas locus, which is absent in all other Bordetella species, has a significantly lower GC-content than the genome-wide average, suggesting acquisition of this locus via horizontal gene transfer from a currently unknown source. The CRISPR array is transcribed and processed into mature CRISPR RNAs (crRNA), some of which have homology to prophages found in closely related species B. hinzii. CONCLUSIONS: Expression of the CRISPR-Cas system and processing of crRNAs with perfect homology to prophages present in closely related species, but absent in that containing this CRISPR-Cas system, suggest it provides protection against phage predation. The 3,117-bp cas9 endonuclease gene from this novel CRISPR-Cas system is 990 bp smaller than that of Streptococcus pyogenes, the 4,017-bp allele currently used for genome editing, and which may make it a useful tool in various CRISPR-Cas technologies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2028-9) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4624362
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-46243622015-10-29 A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease Ivanov, Yury V. Shariat, Nikki Register, Karen B. Linz, Bodo Rivera, Israel Hu, Kai Dudley, Edward G. Harvill, Eric T. BMC Genomics Research Article BACKGROUND: Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR. METHODS: The CRISPR-Cas system has been identified using Basic Local Alignment Search Tool (BLAST) against other sequenced and annotated genomes and confirmed via CRISPRfinder program. Using Polymerase Chain Reactions (PCR) and Sanger DNA sequencing, we discovered CRISPRs in additional bacterial isolates of the same species of Bordetella. Transcriptional activity and processing of the CRISPR have been assessed via RT-PCR. RESULTS: Here we describe a novel Type II-C CRISPR and its associated genes—cas1, cas2, and cas9—in several isolates of a newly discovered Bordetella species. The CRISPR-cas locus, which is absent in all other Bordetella species, has a significantly lower GC-content than the genome-wide average, suggesting acquisition of this locus via horizontal gene transfer from a currently unknown source. The CRISPR array is transcribed and processed into mature CRISPR RNAs (crRNA), some of which have homology to prophages found in closely related species B. hinzii. CONCLUSIONS: Expression of the CRISPR-Cas system and processing of crRNAs with perfect homology to prophages present in closely related species, but absent in that containing this CRISPR-Cas system, suggest it provides protection against phage predation. The 3,117-bp cas9 endonuclease gene from this novel CRISPR-Cas system is 990 bp smaller than that of Streptococcus pyogenes, the 4,017-bp allele currently used for genome editing, and which may make it a useful tool in various CRISPR-Cas technologies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-2028-9) contains supplementary material, which is available to authorized users. BioMed Central 2015-10-26 /pmc/articles/PMC4624362/ /pubmed/26502932 http://dx.doi.org/10.1186/s12864-015-2028-9 Text en © Ivanov et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Ivanov, Yury V.
Shariat, Nikki
Register, Karen B.
Linz, Bodo
Rivera, Israel
Hu, Kai
Dudley, Edward G.
Harvill, Eric T.
A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title_full A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title_fullStr A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title_full_unstemmed A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title_short A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease
title_sort newly discovered bordetella species carries a transcriptionally active crispr-cas with a small cas9 endonuclease
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4624362/
https://www.ncbi.nlm.nih.gov/pubmed/26502932
http://dx.doi.org/10.1186/s12864-015-2028-9
work_keys_str_mv AT ivanovyuryv anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT shariatnikki anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT registerkarenb anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT linzbodo anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT riveraisrael anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT hukai anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT dudleyedwardg anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT harvillerict anewlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT ivanovyuryv newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT shariatnikki newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT registerkarenb newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT linzbodo newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT riveraisrael newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT hukai newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT dudleyedwardg newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease
AT harvillerict newlydiscoveredbordetellaspeciescarriesatranscriptionallyactivecrisprcaswithasmallcas9endonuclease