Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems.
Average rating
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Star rating
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Authors
Fonfara, InesLe Rhun, Anaïs
Chylinski, Krzysztof
Makarova, Kira S
Lécrivain, Anne-Laure
Bzdrenga, Janek
Koonin, Eugene V
Charpentier, Emmanuelle
Issue Date
2013-11-22
Metadata
Show full item recordAbstract
The CRISPR-Cas-derived RNA-guided Cas9 endonuclease is the key element of an emerging promising technology for genome engineering in a broad range of cells and organisms. The DNA-targeting mechanism of the type II CRISPR-Cas system involves maturation of tracrRNA:crRNA duplex (dual-RNA), which directs Cas9 to cleave invading DNA in a sequence-specific manner, dependent on the presence of a Protospacer Adjacent Motif (PAM) on the target. We show that evolution of dual-RNA and Cas9 in bacteria produced remarkable sequence diversity. We selected eight representatives of phylogenetically defined type II CRISPR-Cas groups to analyze possible coevolution of Cas9 and dual-RNA. We demonstrate that these two components are interchangeable only between closely related type II systems when the PAM sequence is adjusted to the investigated Cas9 protein. Comparison of the taxonomy of bacterial species that harbor type II CRISPR-Cas systems with the Cas9 phylogeny corroborates horizontal transfer of the CRISPR-Cas loci. The reported collection of dual-RNA:Cas9 with associated PAMs expands the possibilities for multiplex genome editing and could provide means to improve the specificity of the RNA-programmable Cas9 tool.Citation
Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems. 2013: Nucleic Acids Res.Affiliation
The Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå Centre for Microbial Research (UCMR), Department of Molecular Biology, Umeå University, Umeå S-90187, Sweden, Helmholtz Centre for Infection Research, Department of Regulation in Infection Biology, Braunschweig D-38124, Germany, Deptartment of Biochemistry and Cell Biology, Max F. Perutz Laboratories, University of Vienna, Vienna A-1030, Austria, National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, MD 20894, USA and Hannover Medical School, Hannover D-30625, Germany.Journal
Nucleic acids researchPubMed ID
24270795Type
ArticleISSN
1362-4962ae974a485f413a2113503eed53cd6c53
10.1093/nar/gkt1074
Scopus Count
The following license files are associated with this item:
Related articles
- The tracrRNA and Cas9 families of type II CRISPR-Cas immunity systems.
- Authors: Chylinski K, Le Rhun A, Charpentier E
- Issue date: 2013 May
- crRNA and tracrRNA guide Cas9-mediated DNA interference in Streptococcus thermophilus.
- Authors: Karvelis T, Gasiunas G, Miksys A, Barrangou R, Horvath P, Siksnys V
- Issue date: 2013 May
- The CRISPR-associated DNA-cleaving enzyme Cpf1 also processes precursor CRISPR RNA.
- Authors: Fonfara I, Richter H, Bratovič M, Le Rhun A, Charpentier E
- Issue date: 2016 Apr 28
- DNA interrogation by the CRISPR RNA-guided endonuclease Cas9.
- Authors: Sternberg SH, Redding S, Jinek M, Greene EC, Doudna JA
- Issue date: 2014 Mar 6
- Classification and evolution of type II CRISPR-Cas systems.
- Authors: Chylinski K, Makarova KS, Charpentier E, Koonin EV
- Issue date: 2014 Jun