Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials

被引：638

作者：

Bikard, David ^{[1
]}

Euler, Chad W. ^{[2
]}

Jiang, Wenyan ^{[1
]}

Nussenzweig, Philip M. ^{[1
]}

Goldberg, Gregory W. ^{[1
]}

Duportet, Xavier ^{[3
,4
]}

Fischetti, Vincent A. ^{[2
]}

Marraffini, Luciano A. ^{[1
]}

机构：

[1] Rockefeller Univ, Bacteriol Lab, New York, NY 10021 USA

[2] Rockefeller Univ, Lab Bacterial Pathogenesis & Immunol, New York, NY 10021 USA

[3] INRIA Paris Rocquencourt, Rocquencourt, France

[4] MIT, Synthet Biol Ctr, Dept Biol Engn, Cambridge, MA 02139 USA

来源：

NATURE BIOTECHNOLOGY | 2014年 / 32卷 / 11期

关键词：

STAPHYLOCOCCUS-AUREUS; GENOME SEQUENCE; RNA; GENE; EXPRESSION; ENCODES;

D O I：

10.1038/nbt.3043

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Antibiotics target conserved bacterial cellular pathways or growth functions and therefore cannot selectively kill specific members of a complex microbial population. Here, we develop programmable, sequence-specific antimicrobials using the RNA-guided nuclease Cas9 (refs.1,2) delivered by a bacteriophage. We show that Cas9, reprogrammed to target virulence genes, kills virulent, but not avirulent, Staphylococcus aureus. Reprogramming the nuclease to target antibiotic resistance genes destroys staphylococcal plasmids that harbor antibiotic resistance genes(3,4) and immunizes avirulent staphylococci to prevent the spread of plasmid-borne resistance genes. We also show that CRISPR-Cas9 antimicrobials function in vivo to kill S. aureus in a mouse skin colonization model. This technology creates opportunities to manipulate complex bacterial populations in a sequence-specific manner.

引用

页码：1146 / 1150

页数：5

共 26 条

[1]

Agarwal R, 2013, THER DELIV, V4, P705, DOI [10.4155/TDE.13.37, 10.4155/tde.13.37]

[2] Genome sequence of Staphylococcus aureus strain newman and comparative analysis of staphylococcal genomes:: Polymorphism and evolution of two major pathogenicity islands [J].

Baba, Tadashi ;

Bae, Taeok ;

Schneewind, Olaf ;

Takeuchi, Fumihiko ;

Hiramatsu, Keiichi .

JOURNAL OF BACTERIOLOGY, 2008, 190 (01) :300-310

[3] Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system [J].

Bikard, David ;

Jiang, Wenyan ;

Samai, Poulami ;

Hochschild, Ann ;

Zhang, Feng ;

Marraffini, Luciano A. .

NUCLEIC ACIDS RESEARCH, 2013, 41 (15) :7429-7437

[4] CRISPR Interference Can Prevent Natural Transformation and Virulence Acquisition during In Vivo Bacterial Infection [J].

Bikard, David ;

Hatoum-Aslan, Asma ;

Mucida, Daniel ;

Marraffini, Luciano A. .

CELL HOST & MICROBE, 2012, 12 (02) :177-186

[5] Novel cassette-based shuttle vector system for gram-positive bacteria [J].

Charpentier, E ;

Anton, AI ;

Barry, P ;

Alfonso, B ;

Fang, Y ;

Novick, RP .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (10) :6076-6085

[6] CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III [J].

Deltcheva, Elitza ;

Chylinski, Krzysztof ;

Sharma, Cynthia M. ;

Gonzales, Karine ;

Chao, Yanjie ;

Pirzada, Zaid A. ;

Eckert, Maria R. ;

Vogel, Joerg ;

Charpentier, Emmanuelle .

NATURE, 2011, 471 (7340) :602-+

[7] The Pervasive Effects of an Antibiotic on the Human Gut Microbiota, as Revealed by Deep 16S rRNA Sequencing [J].

Dethlefsen, Les ;

Huse, Sue ;

Sogin, Mitchell L. ;

Relman, David A. .

PLOS BIOLOGY, 2008, 6 (11) :2383-2400

[8] Complete genome sequence of USA300, an epidemic clone of community-acquired meticillin-resistant Staphylococcus aureus [J].

Diep, BA ;

Gill, SR ;

Chang, RF ;

Phan, TH ;

Chen, JH ;

Davidson, MG ;

Lin, F ;

Lin, J ;

Carleton, HA ;

Mongodin, EF ;

Sensabaugh, GF ;

Perdreau-Remington, F .

LANCET, 2006, 367 (9512) :731-739

[9] Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria [J].

Gasiunas, Giedrius ;

Barrangou, Rodolphe ;

Horvath, Philippe ;

Siksnys, Virginijus .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2012, 109 (39) :E2579-E2586

[10] Programmable Removal of Bacterial Strains by Use of Genome-Targeting CRISPR-Cas Systems [J].

Gomaa, Ahmed A. ;

Klumpe, Heidi E. ;

Luo, Michelle L. ;

Selle, Kurt ;

Barrangou, Rodolphe ;

Beisel, Chase L. .

MBIO, 2014, 5 (01)

← 1 2 3 →