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The DOI matches its Crossref record; 10 of the 10 references it cites were found in a registry or an index.

Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems

Fonfara, Ines, Le Rhun, Anaïs, Chylinski, Krzysztof, Makarova, Kira S., Lécrivain, Anne-Laure, Bzdrenga, Janek, Koonin, Eugene V., Charpentier, Emmanuelle·2014·Nucleic Acids Research
DOI 10.1093/nar/gkt1074PMC PMC3936727
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Abstract

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.

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  • Kim et al. determined that CjCas9 could generally cleave target sites with the PAM site 5′-NNNNRYAC (where Y = pyrimidine and R = purine).

    “by testing a variety of PAM sites, Kim et al. determined that CjCas9 could generally cleave target sites with the PAM site 5′-NNNNRYAC (where Y = pyrimidine and R = purine) [72,73].”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [73]

  • Fonfara et al. reported the optimal in vitro PAM site for CjCas9 to be 5′-NNNNACA.

    “Fonfara et al. previously reported the optimal in vitro PAM site for CjCas9 to be 5′-NNNNACA”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [73]

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Metadata

Authors
Fonfara, I., Le Rhun, A., Chylinski, K., Makarova, K. S., Lécrivain, A.-L., Bzdrenga, J., Koonin, E. V., & Charpentier, E.
Affiliations
The Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå Centre for Microbial Research (UCMR), Department of Molecular Biology, Umeå University, Umeå S-90187, Sweden, 2Helmholtz Centre for Infection Research, Department of Regulation in Infection Biology, Braunschweig D-38124, Germany, 3Deptartment of Biochemistry and Cell Biology, Max F. Perutz Laboratories, University of Vienna, Vien
Venue
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Published
2013-11-21
Volume
42
Issue
4
Pages
2577-2590
ISSN
  • 1362-4962
  • 0305-1048
DOI
10.1093/nar/gkt1074
PMC
PMC3936727
Cited by
328
References
10
Language
en
Licence
creativecommons.org/licenses/by/3.0/ (vor, 1085 d)

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