Cargando…
High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon
CRISPR-Cas systems allow bacteria and archaea to acquire sequence-specific immunity against selfish genetic elements such as viruses and plasmids, by specific degradation of invader DNA or RNA. However, this involves the risk of autoimmunity if immune memory against host DNA is mistakenly acquired....
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435918/ https://www.ncbi.nlm.nih.gov/pubmed/28334774 http://dx.doi.org/10.1093/nar/gkx150 |
_version_ | 1783237306091569152 |
---|---|
author | Stachler, Aris-Edda Turgeman-Grott, Israela Shtifman-Segal, Ella Allers, Thorsten Marchfelder, Anita Gophna, Uri |
author_facet | Stachler, Aris-Edda Turgeman-Grott, Israela Shtifman-Segal, Ella Allers, Thorsten Marchfelder, Anita Gophna, Uri |
author_sort | Stachler, Aris-Edda |
collection | PubMed |
description | CRISPR-Cas systems allow bacteria and archaea to acquire sequence-specific immunity against selfish genetic elements such as viruses and plasmids, by specific degradation of invader DNA or RNA. However, this involves the risk of autoimmunity if immune memory against host DNA is mistakenly acquired. Such autoimmunity has been shown to be highly toxic in several bacteria and is believed to be one of the major costs of maintaining these defense systems. Here we generated an experimental system in which a non-essential gene, required for pigment production and the reddish colony color, is targeted by the CRISPR-Cas I-B system of the halophilic archaeon Haloferax volcanii. We show that under native conditions, where both the self-targeting and native crRNAs are expressed, self-targeting by CRISPR-Cas causes no reduction in transformation efficiency of the plasmid encoding the self-targeting crRNA. Furthermore, under such conditions, no effect on organismal growth rate or loss of the reddish colony phenotype due to mutations in the targeted region could be observed. In contrast, in cells deleted for the pre-crRNA processing gene cas6, where only the self-targeting crRNA exists as mature crRNA, self-targeting leads to moderate toxicity and the emergence of deletion mutants. Sequencing of the deletions caused by CRISPR-Cas self targeting indicated DNA repair via microhomology-mediated end joining. |
format | Online Article Text |
id | pubmed-5435918 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-54359182017-05-22 High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon Stachler, Aris-Edda Turgeman-Grott, Israela Shtifman-Segal, Ella Allers, Thorsten Marchfelder, Anita Gophna, Uri Nucleic Acids Res Genome Integrity, Repair and Replication CRISPR-Cas systems allow bacteria and archaea to acquire sequence-specific immunity against selfish genetic elements such as viruses and plasmids, by specific degradation of invader DNA or RNA. However, this involves the risk of autoimmunity if immune memory against host DNA is mistakenly acquired. Such autoimmunity has been shown to be highly toxic in several bacteria and is believed to be one of the major costs of maintaining these defense systems. Here we generated an experimental system in which a non-essential gene, required for pigment production and the reddish colony color, is targeted by the CRISPR-Cas I-B system of the halophilic archaeon Haloferax volcanii. We show that under native conditions, where both the self-targeting and native crRNAs are expressed, self-targeting by CRISPR-Cas causes no reduction in transformation efficiency of the plasmid encoding the self-targeting crRNA. Furthermore, under such conditions, no effect on organismal growth rate or loss of the reddish colony phenotype due to mutations in the targeted region could be observed. In contrast, in cells deleted for the pre-crRNA processing gene cas6, where only the self-targeting crRNA exists as mature crRNA, self-targeting leads to moderate toxicity and the emergence of deletion mutants. Sequencing of the deletions caused by CRISPR-Cas self targeting indicated DNA repair via microhomology-mediated end joining. Oxford University Press 2017-05-19 2017-03-02 /pmc/articles/PMC5435918/ /pubmed/28334774 http://dx.doi.org/10.1093/nar/gkx150 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Genome Integrity, Repair and Replication Stachler, Aris-Edda Turgeman-Grott, Israela Shtifman-Segal, Ella Allers, Thorsten Marchfelder, Anita Gophna, Uri High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title | High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title_full | High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title_fullStr | High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title_full_unstemmed | High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title_short | High tolerance to self-targeting of the genome by the endogenous CRISPR-Cas system in an archaeon |
title_sort | high tolerance to self-targeting of the genome by the endogenous crispr-cas system in an archaeon |
topic | Genome Integrity, Repair and Replication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435918/ https://www.ncbi.nlm.nih.gov/pubmed/28334774 http://dx.doi.org/10.1093/nar/gkx150 |
work_keys_str_mv | AT stachlerarisedda hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon AT turgemangrottisraela hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon AT shtifmansegalella hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon AT allersthorsten hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon AT marchfelderanita hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon AT gophnauri hightolerancetoselftargetingofthegenomebytheendogenouscrisprcassysteminanarchaeon |