Open Access

Science, volume 337, issue 6096, pages 816-821

A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity

Jinek Martin ^{1, 2}

Chyliński Krzysztof ^{3, 4}

FONFARA INES ⁴

Hauer Michael ²

Doudna Jennifer A. ^{1, 2, 5, 6}

Charpentier Emmanuelle ⁴

Hide authors affiliations Show authors affiliations: 6 affiliations

Howard Hughes Medical Institute (HHMI), University of California, Berkeley, CA 94720, USA. |

Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA. |

Max F. Perutz Laboratories (MFPL), University of Vienna, A-1030 Vienna, Austria. |

⁴

The Laboratory for Molecular Infection Medicine Sweden, Umeå Centre for Microbial Research, Department of Molecular Biology, Umeå University, S-90187 Umeå, Sweden. |

⁵

Department of Chemistry, University of California, Berkeley, CA 94720, USA. |

⁶

Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. |

Publication type: Journal Article

Publication date: 2012-06-28

American Association for the Advancement of Science (AAAS)

Journal: Science

Quartile SCImago

Quartile WOS

Impact factor: 56.9

ISSN: 00368075, 10959203

DOI: 10.1126/science.1225829

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PubMed ID: 22745249

Multidisciplinary

Abstract

Ditching Invading DNA Bacteria and archaea protect themselves from invasive foreign nucleic acids through an RNA-mediated adaptive immune system called CRISPR (clustered regularly interspaced short palindromic repeats)/CRISPR-associated (Cas). Jinek et al. (p. 816, published online 28 June; see the Perspective by Brouns) found that for the type II CRISPR/Cas system, the CRISPR RNA (crRNA) as well as the trans-activating crRNA—which is known to be involved in the pre-crRNA processing—were both required to direct the Cas9 endonuclease to cleave the invading target DNA. Furthermore, engineered RNA molecules were able to program the Cas9 endonuclease to cleave specific DNA sequences to generate double-stranded DNA breaks. A prokaryotic RNA–directed targeting system can be designed to cleave any DNA sequence. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.

By date By citations

Citations by journals

	50 100 150 200 250 300 350
Methods in Molecular Biology	Methods in Molecular Biology, 316, 2.71% Methods in Molecular Biology 316 publications, 2.71%
Scientific Reports	Scientific Reports, 257, 2.21% Scientific Reports 257 publications, 2.21%
Nucleic Acids Research	Nucleic Acids Research, 242, 2.08% Nucleic Acids Research 242 publications, 2.08%
International Journal of Molecular Sciences	International Journal of Molecular Sciences, 215, 1.85% International Journal of Molecular Sciences 215 publications, 1.85%
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PLoS ONE	PLoS ONE, 142, 1.22% PLoS ONE 142 publications, 1.22%
Frontiers in Plant Science	Frontiers in Plant Science, 122, 1.05% Frontiers in Plant Science 122 publications, 1.05%
ACS Synthetic Biology	ACS Synthetic Biology, 103, 0.88% ACS Synthetic Biology 103 publications, 0.88%
Nature Biotechnology	Nature Biotechnology, 100, 0.86% Nature Biotechnology 100 publications, 0.86%
The CRISPR Journal	The CRISPR Journal, 89, 0.76% The CRISPR Journal 89 publications, 0.76%
Molecular Cell	Molecular Cell, 88, 0.76% Molecular Cell 88 publications, 0.76%
Proceedings of the National Academy of Sciences of the United States of America	Proceedings of the National Academy of Sciences of the United States of America, 85, 0.73% Proceedings of the National Academy of Sciences of the United States of America 85 publications, 0.73%
Nature	Nature, 77, 0.66% Nature 77 publications, 0.66%
Genes	Genes, 71, 0.61% Genes 71 publications, 0.61%
Molecular Therapy	Molecular Therapy, 70, 0.6% Molecular Therapy 70 publications, 0.6%
Cells	Cells, 68, 0.58% Cells 68 publications, 0.58%
eLife	eLife, 65, 0.56% eLife 65 publications, 0.56%
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Cell	Cell, 63, 0.54% Cell 63 publications, 0.54%
Molecular Therapy - Nucleic Acids	Molecular Therapy - Nucleic Acids, 61, 0.52% Molecular Therapy - Nucleic Acids 61 publications, 0.52%
Science	Science, 61, 0.52% Science 61 publications, 0.52%
Frontiers in Genetics	Frontiers in Genetics, 57, 0.49% Frontiers in Genetics 57 publications, 0.49%
Frontiers in Bioengineering and Biotechnology	Frontiers in Bioengineering and Biotechnology, 55, 0.47% Frontiers in Bioengineering and Biotechnology 55 publications, 0.47%
Plant Biotechnology Journal	Plant Biotechnology Journal, 55, 0.47% Plant Biotechnology Journal 55 publications, 0.47%
Methods	Methods, 54, 0.46% Methods 54 publications, 0.46%
Frontiers in Microbiology	Frontiers in Microbiology, 48, 0.41% Frontiers in Microbiology 48 publications, 0.41%
Nature Protocols	Nature Protocols, 47, 0.4% Nature Protocols 47 publications, 0.4%
Progress in Molecular Biology and Translational Science	Progress in Molecular Biology and Translational Science, 47, 0.4% Progress in Molecular Biology and Translational Science 47 publications, 0.4%
Genetics	Genetics, 47, 0.4% Genetics 47 publications, 0.4%
	50 100 150 200 250 300 350

Citations by publishers

	500 1000 1500 2000 2500
Springer Nature	Springer Nature, 2480, 21.29% Springer Nature 2480 publications, 21.29%
Elsevier	Elsevier, 2052, 17.61% Elsevier 2052 publications, 17.61%
Wiley	Wiley, 824, 7.07% Wiley 824 publications, 7.07%
Multidisciplinary Digital Publishing Institute (MDPI)	Multidisciplinary Digital Publishing Institute (MDPI), 719, 6.17% Multidisciplinary Digital Publishing Institute (MDPI) 719 publications, 6.17%
Frontiers Media S.A.	Frontiers Media S.A., 559, 4.8% Frontiers Media S.A. 559 publications, 4.8%
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American Chemical Society (ACS)	American Chemical Society (ACS), 333, 2.86% American Chemical Society (ACS) 333 publications, 2.86%
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Mary Ann Liebert	Mary Ann Liebert, 163, 1.4% Mary Ann Liebert 163 publications, 1.4%
American Society for Microbiology	American Society for Microbiology, 124, 1.06% American Society for Microbiology 124 publications, 1.06%
American Association for the Advancement of Science (AAAS)	American Association for the Advancement of Science (AAAS), 107, 0.92% American Association for the Advancement of Science (AAAS) 107 publications, 0.92%
Royal Society of Chemistry (RSC)	Royal Society of Chemistry (RSC), 103, 0.88% Royal Society of Chemistry (RSC) 103 publications, 0.88%
Genetics Society of America	Genetics Society of America, 87, 0.75% Genetics Society of America 87 publications, 0.75%
Proceedings of the National Academy of Sciences (PNAS)	Proceedings of the National Academy of Sciences (PNAS), 85, 0.73% Proceedings of the National Academy of Sciences (PNAS) 85 publications, 0.73%
The Company of Biologists	The Company of Biologists, 68, 0.58% The Company of Biologists 68 publications, 0.58%
Cold Spring Harbor Laboratory	Cold Spring Harbor Laboratory, 66, 0.57% Cold Spring Harbor Laboratory 66 publications, 0.57%
eLife Sciences Publications	eLife Sciences Publications, 65, 0.56% eLife Sciences Publications 65 publications, 0.56%
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Wolters Kluwer Health	Wolters Kluwer Health, 47, 0.4% Wolters Kluwer Health 47 publications, 0.4%
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Pleiades Publishing	Pleiades Publishing, 44, 0.38% Pleiades Publishing 44 publications, 0.38%
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KeAi Communications Co.	KeAi Communications Co., 37, 0.32% KeAi Communications Co. 37 publications, 0.32%
Bentham Science	Bentham Science, 35, 0.3% Bentham Science 35 publications, 0.3%
American Society for Biochemistry and Molecular Biology	American Society for Biochemistry and Molecular Biology, 34, 0.29% American Society for Biochemistry and Molecular Biology 34 publications, 0.29%
Hindawi Limited	Hindawi Limited, 27, 0.23% Hindawi Limited 27 publications, 0.23%
The Royal Society	The Royal Society, 22, 0.19% The Royal Society 22 publications, 0.19%
EMBO press	EMBO press, 22, 0.19% EMBO press 22 publications, 0.19%
	500 1000 1500 2000 2500

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GOST Copy

Jinek M. et al. A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity // Science. 2012. Vol. 337. No. 6096. pp. 816-821.

GOST all authors (up to 50) Copy

Jinek M., Chyliński K., FONFARA I., Hauer M., Doudna J. A., Charpentier E. A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity // Science. 2012. Vol. 337. No. 6096. pp. 816-821.

RIS |

Cite this

RIS Copy

TY - JOUR

DO - 10.1126/science.1225829

UR - https://doi.org/10.1126%2Fscience.1225829

TI - A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity

T2 - Science

AU - Jinek, Martin

AU - Chyliński, Krzysztof

AU - FONFARA, INES

AU - Hauer, Michael

AU - Doudna, Jennifer A.

AU - Charpentier, Emmanuelle

PY - 2012

DA - 2012/06/28 00:00:00

PB - American Association for the Advancement of Science (AAAS)

SP - 816-821

IS - 6096

VL - 337

PMID - 22745249

SN - 0036-8075

SN - 1095-9203

ER -

BibTex |

Cite this

BibTex Copy

@article{2012_Jinek,

author = {Martin Jinek and Krzysztof Chyliński and INES FONFARA and Michael Hauer and Jennifer A. Doudna and Emmanuelle Charpentier},

title = {A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity},

journal = {Science},

year = {2012},

volume = {337},

publisher = {American Association for the Advancement of Science (AAAS)},

month = {jun},

url = {https://doi.org/10.1126%2Fscience.1225829},

number = {6096},

pages = {816--821},

doi = {10.1126/science.1225829}

}

MLA

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MLA Copy

Jinek, Martin, et al. “A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity.” Science, vol. 337, no. 6096, Jun. 2012, pp. 816-821. https://doi.org/10.1126%2Fscience.1225829.

Found error?

Publisher

American Association for the Advancement of Science (AAAS)

Journal

Science

Quartile SCImago

Quartile WOS

Impact factor

56.9

ISSN

00368075 (Print)
10959203 (Electronic)