Key Points
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Recently, arrays of clustered, regularly interspaced short palindromic repeats (CRISPRs) have been implicated in a novel genetic interference pathway that limits phage infection and plasmid conjugation. CRISPR loci keep a record of past infections to provide bacteria and archaea with a 'genetic memory' that directs the rejection of invader DNA molecules; therefore these loci constitute an adaptive immune system.
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CRISPRs (which are approximately 25–50 nucleotides long) are separated by similarly short sequences called spacers that match bacteriophage or plasmid sequences and specify the targets of interference. CRISPR-associated (cas) genes, a set of conserved genes that are associated with these loci, are usually present on one or the other side of the array.
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CRISPR loci are transcribed as a long precursor that is processed by Cas proteins within the repeat sequences to generate small CRISPR RNAs (crRNAs). The crRNAs serve as guides for target recognition during CRISPR interference. crRNA–Cas ribonucleoprotein complexes seem to generally target invading DNA sequences during interference but may also target RNAs in some species.
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Upon bacteriophage challenge of a CRISPR-containing bacterial population, mutants resistant to the infection arise through the incorporation of additional spacer sequences derived from the challenging phage. This allows the bacteria to evolve rapidly and adapt to the viruses in the environment.
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Bacteriophages and plasmids can mobilize foreign genetic material between cells, a process known as horizontal gene transfer (HGT), which is a fundamental source of genetic variability for the evolution of bacteria and archaea. CRISPR interference prevents phage infection and plasmid conjugation and therefore constitutes a natural barrier to HGT. In addition, bacteriophages constantly mutate to evade CRISPR defence. Therefore, CRISPR interference has an important role in the evolution of microbial communities.
Abstract
Sequence-directed genetic interference pathways control gene expression and preserve genome integrity in all kingdoms of life. The importance of such pathways is highlighted by the extensive study of RNA interference (RNAi) and related processes in eukaryotes. In many bacteria and most archaea, clustered, regularly interspaced short palindromic repeats (CRISPRs) are involved in a more recently discovered interference pathway that protects cells from bacteriophages and conjugative plasmids. CRISPR sequences provide an adaptive, heritable record of past infections and express CRISPR RNAs — small RNAs that target invasive nucleic acids. Here, we review the mechanisms of CRISPR interference and its roles in microbial physiology and evolution. We also discuss potential applications of this novel interference pathway.
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References
Nakamura, Y., Itoh, T., Matsuda, H. & Gojobori, T. Biased biological functions of horizontally transferred genes in prokaryotic genomes. Nature Genet. 36, 760–766 (2004).
Thomas, C. M. & Nielsen, K. M. Mechanisms of, and barriers to, horizontal gene transfer between bacteria. Nature Rev. Microbiol. 3, 711–721 (2005).
Ishino, Y., Shinagawa, H., Makino, K., Amemura, M. & Nakata, A. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J. Bacteriol. 169, 5429–5433 (1987).
Nakata, A., Amemura, M. & Makino, K. Unusual nucleotide arrangement with repeated sequences in the Escherichia coli K-12 chromosome. J. Bacteriol. 171, 3553–3556 (1989).
Hermans, P. W. et al. Insertion element IS987 from Mycobacterium bovis BCG is located in a hot-spot integration region for insertion elements in Mycobacterium tuberculosis complex strains. Infect. Immun. 59, 2695–2705 (1991).
Mojica, F. J., Ferrer, C., Juez, G. & Rodriguez-Valera, F. Long stretches of short tandem repeats are present in the largest replicons of the Archaea Haloferax mediterranei and Haloferax volcanii and could be involved in replicon partitioning. Mol. Microbiol. 17, 85–93 (1995).
Masepohl, B., Gorlitz, K. & Bohme, H. Long tandemly repeated repetitive (LTRR) sequences in the filamentous cyanobacterium Anabaena sp. PCC 7120. Biochim. Biophys. Acta 1307, 26–30 (1996).
Hoe, N. et al. Rapid molecular genetic subtyping of serotype M1 group A Streptococcus strains. Emerg. Infect. Dis. 5, 254–263 (1999).
Bult, C. J. et al. Complete genome sequence of the methanogenic archaeon, Methanococcus jannaschii. Science 273, 1058–1073 (1996).
Klenk, H. P. et al. The complete genome sequence of the hyperthermophilic, sulphate-reducing archaeon Archaeoglobus fulgidus. Nature 390, 364–370 (1997).
Nelson, K. E. et al. Evidence for lateral gene transfer between Archaea and Bacteria from genome sequence of Thermotoga maritima. Nature 399, 323–329 (1999).
Sensen, C. W. et al. Completing the sequence of the Sulfolobus solfataricus P2 genome. Extremophiles 2, 305–312 (1998).
Kawarabayasi, Y. et al. Complete genome sequence of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1. DNA Res. 6, 83–101 (1999).
Kawarabayasi, Y. et al. Complete sequence and gene organization of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3. DNA Res. 5, 55–76 (1998).
Mojica, F. J., Diez-Villasenor, C., Soria, E. & Juez, G. Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol. Microbiol. 36, 244–246 (2000).
Jansen, R., Embden, J. D., Gaastra, W. & Schouls, L. M. Identification of genes that are associated with DNA repeats in prokaryotes. Mol. Microbiol. 43, 1565–1575 (2002). This paper noted the existence of cas protein-coding genes associated with CRISPR loci.
Bolotin, A., Quinquis, B., Sorokin, A. & Ehrlich, S. D. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology 151, 2551–2561 (2005). This paper, along with references 18 and 19, documented the sequence matches between CRISPR spacers and phage or plasmid sequences, thereby suggesting a role for CRISPR loci in defence against invasive DNA.
Mojica, F. J., Diez-Villasenor, C., Garcia-Martinez, J. & Soria, E. Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J. Mol. Evol. 60, 174–182 (2005).
Pourcel, C., Salvignol, G. & Vergnaud, G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology 151, 653–663 (2005).
Lillestøl, R. K., Redder, P., Garrett, R. A. & Brugger, K. A putative viral defence mechanism in archaeal cells. Archaea 2, 59–72 (2006).
Makarova, K. S., Grishin, N. V., Shabalina, S. A., Wolf, Y. I. & Koonin, E. V. A putative RNA-interference-based immune system in prokaryotes: computational analysis of the predicted enzymatic machinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biol. Direct 1, 7 (2006).
Barrangou, R. et al. CRISPR provides acquired resistance against viruses in prokaryotes. Science 315, 1709–1712 (2007). This study provided the first experimental demonstration of CRISPR interference, documented the acquisition of novel spacers in response to phage infection and showed that cas gene function is essential for CRISPR function in defence.
Marraffini, L. A. & Sontheimer, E. J. CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science 322, 1843–1845 (2008). This paper showed that CRISPR interference also blocks conjugation, and demonstrated that DNA rather than RNA is the molecular target of the pathway in S. epidermidis.
Grissa, I., Vergnaud, G. & Pourcel, C. The CRISPRdb database and tools to display CRISPRs and to generate dictionaries of spacers and repeats. BMC Bioinformatics 8, 172 (2007).
Kunin, V., Sorek, R. & Hugenholtz, P. Evolutionary conservation of sequence and secondary structures in CRISPR repeats. Genome Biol. 8, R61 (2007).
Andersson, A. F. & Banfield, J. F. Virus population dynamics and acquired virus resistance in natural microbial communities. Science 320, 1047–1050 (2008). This study used metagenomics to profile CRISPR spacer composition and phage sequences in a spatially and temporally resolved fashion to analyse host–pathogen interactions in a natural microbial population.
Heidelberg, J. F., Nelson, W. C., Schoenfeld, T. & Bhaya, D. Germ warfare in a microbial mat community: CRISPRs provide insights into the co-evolution of host and viral genomes. PLoS ONE 4, e4169 (2009).
Haft, D. H., Selengut, J., Mongodin, E. F. & Nelson, K. E. A guild of 45 CRISPR-associated (Cas) protein families and multiple CRISPR/Cas subtypes exist in prokaryotic genomes. PLoS Comput. Biol. 1, e60 (2005). This work, along with reference 21, examined CRISPR and cas phylogenetics and established the existence of distinct subtypes of CRISPR– cas loci and multiple families of Cas proteins.
Shah, S. A., Hansen, N. R. & Garrett, R. A. Distribution of CRISPR spacer matches in viruses and plasmids of crenarchaeal acidothermophiles and implications for their inhibitory mechanism. Biochem. Soc. Trans. 37, 23–28 (2009).
Godde, J. S. & Bickerton, A. The repetitive DNA elements called CRISPRs and their associated genes: evidence of horizontal transfer among prokaryotes. J. Mol. Evol. 62, 718–729 (2006).
Makarova, K. S., Aravind, L., Grishin, N. V., Rogozin, I. B. & Koonin, E. V. A DNA repair system specific for thermophilic Archaea and Bacteria predicted by genomic context analysis. Nucleic Acids Res. 30, 482–496 (2002).
Portillo, M. C. & Gonzalez, J. M. CRISPR elements in the Thermococcales: evidence for associated horizontal gene transfer in Pyrococcus furiosus. J. Appl. Genet. 50, 421–430 (2009).
van der Oost, J., Jore, M. M., Westra, E. R., Lundgren, M. & Brouns, S. J. CRISPR-based adaptive and heritable immunity in prokaryotes. Trends Biochem. Sci. 34, 401–407 (2009).
van der Ploeg, J. R. Analysis of CRISPR in Streptococcus mutans suggests frequent occurrence of acquired immunity against infection by M102-like bacteriophages. Microbiology 155, 1966–1976 (2009).
Deveau, H. et al. Phage response to CRISPR-encoded resistance in Streptococcus thermophilus. J. Bacteriol. 190, 1390–1400 (2008). This paper, along with reference 40, reported the existence of short, conserved 'CRISPR motifs' or 'protospacer-adjacent motifs' that flank target protospacers.
Horvath, P. et al. Diversity, activity, and evolution of CRISPR loci in Streptococcus thermophilus. J. Bacteriol. 190, 1401–1412 (2008).
Tyson, G. W. & Banfield, J. F. Rapidly evolving CRISPRs implicated in acquired resistance of microorganisms to viruses. Environ. Microbiol. 10, 200–207 (2007).
Brouns, S. J. et al. Small CRISPR RNAs guide antiviral defense in prokaryotes. Science 321, 960–964 (2008). This report, along with reference 49, documented and characterized the crRNA processing pathway; Brouns et al . also proved that crRNAs are essential for CRISPR interference in E. coli , implicated Cas3 as an effector protein in this process and pointed towards DNA as a likely target.
Wiedenheft, B. et al. Structural basis for DNase activity of a conserved protein implicated in CRISPR-mediated genome defense. Structure 17, 904–912 (2009). This study reported the first crystal structure of a protein (Cas1) that is universal among CRISPR– cas loci and documented its nuclease activity.
Mojica, F. J., Diez-Villasenor, C., Garcia-Martinez, J. & Almendros, C. Short motif sequences determine the targets of the prokaryotic CRISPR defence system. Microbiology 155, 733–740 (2009).
Semenova, E., Nagornykh, M., Pyatnitskiy, M., Artamonova, I. I. & Severinov, K. Analysis of CRISPR system function in plant pathogen Xanthomonas oryzae. FEMS Microbiol. Lett. 296, 110–116 (2009).
Tang, T. H. et al. Identification of 86 candidates for small non-messenger RNAs from the archaeon Archaeoglobus fulgidus. Proc. Natl Acad. Sci. USA 99, 7536–7541 (2002).
Tang, T. H. et al. Identification of novel non-coding RNAs as potential antisense regulators in the archaeon Sulfolobus solfataricus. Mol. Microbiol. 55, 469–481 (2005).
Hale, C., Kleppe, K., Terns, R. M. & Terns, M. P. Prokaryotic silencing (psi)RNAs in Pyrococcus furiosus. RNA 14, 2572–2579 (2008).
Lillestøl, R. K. et al. CRISPR families of the crenarchaeal genus Sulfolobus: bidirectional transcription and dynamic properties. Mol. Microbiol. 72, 259–272 (2009).
Agari, Y. et al. Transcription profile of Thermus thermophilus CRISPR systems after phage infection. J. Mol. Biol. 395, 270–281 (2010).
Shinkai, A. et al. Transcription activation mediated by a cyclic AMP receptor protein from Thermus thermophilus HB8. J. Bacteriol. 189, 3891–3901 (2007).
Chattoraj, P., Banerjee, A., Biswas, S. & Biswas, I. ClpP of Streptococcus mutans differentially regulates expression of genomic islands, mutacin production and antibiotics tolerance. J. Bacteriol.28 Dec 2009 (doi:10.1128/JB.01350-09).
Carte, J., Wang, R., Li, H., Terns, R. M. & Terns, M. P. Cas6 is an endoribonuclease that generates guide RNAs for invader defense in prokaryotes. Genes Dev. 22, 3489–3496 (2008).
Hale, C. R. et al. RNA-guided RNA cleavage by a CRISPR RNA–Cas protein complex. Cell 139, 945–956 (2009). This paper characterized P. furiosus crRNP components, reported the first high-throughput profiling of a crRNA population and demonstrated crRNA-directed RNA cleavage by a crRNP complex in vitro.
Ebihara, A. et al. Crystal structure of hypothetical protein TTHB192 from Thermus thermophilus HB8 reveals a new protein family with an RNA recognition motif-like domain. Protein Sci. 15, 1494–1499 (2006).
Redder, P. et al. Four newly isolated fuselloviruses from extreme geothermal environments reveal unusual morphologies and a possible interviral recombination mechanism. Environ. Microbiol. 11, 2849–2862 (2009).
Cui, Y. et al. Insight into microevolution of Yersinia pestis by clustered regularly interspaced short palindromic repeats. PLoS ONE 3, e2652 (2008).
Miller, E. S. et al. Bacteriophage T4 genome. Microbiol. Mol. Biol. Rev. 67, 86–156 (2003).
Gill, S. R. et al. Insights on evolution of virulence and resistance from the complete genome analysis of an early methicillin-resistant Staphylococcus aureus strain and a biofilm-producing methicillin-resistant Staphylococcus epidermidis strain. J. Bacteriol. 187, 2426–2438 (2005).
Climo, M. W., Sharma, V. K. & Archer, G. L. Identification and characterization of the origin of conjugative transfer (oriT) and a gene (nes) encoding a single-stranded endonuclease on the staphylococcal plasmid pGO1. J. Bacteriol. 178, 4975–4983 (1996).
Marraffini, L. A. & Sontheimer, E. J. Self versus non-self discrimination during CRISPR RNA-directed immunity. Nature13 Jan 2010 (doi:10.1038/nature08703).
Han, D. & Krauss, G. Characterization of the endonuclease SSO2001 from Sulfolobus solfataricus P2. FEBS Lett. 583, 771–776 (2009).
Furuya, E. Y. & Lowy, F. D. Antimicrobial-resistant bacteria in the community setting. Nature Rev. Microbiol. 4, 36–45 (2006).
Weigel, L. M. et al. Genetic analysis of a high-level vancomycin-resistant isolate of Staphylococcus aureus. Science 302, 1569–1571 (2003).
Diep, B. A. et al. Complete genome sequence of USA300, an epidemic clone of community-acquired meticillin-resistant Staphylococcus aureus. Lancet 367, 731–739 (2006).
Lim, S. M. & Webb, S. A. Nosocomial bacterial infections in intensive care units. I: Organisms and mechanisms of antibiotic resistance. Anaesthesia 60, 887–902 (2005).
Lowy, F. D. Staphylococcus aureus infections. N. Engl. J. Med. 339, 520–532 (1998).
von Eiff, C., Peters, G. & Heilmann, C. Pathogenesis of infections due to coagulase-negative staphylococci. Lancet Infect. Dis. 2, 677–685 (2002).
Brussow, H., Canchaya, C. & Hardt, W. D. Phages and the evolution of bacterial pathogens: from genomic rearrangements to lysogenic conversion. Microbiol. Mol. Biol. Rev. 68, 560–602 (2004).
Banks, D. J., Beres, S. B. & Musser, J. M. The fundamental contribution of phages to GAS evolution, genome diversification and strain emergence. Trends Microbiol. 10, 515–521 (2002).
Hu, X., Van der Auwera, G., Timmery, S., Zhu, L. & Mahillon, J. Distribution, diversity, and potential mobility of extrachromosomal elements related to the Bacillus anthracis pXO1 and pXO2 virulence plasmids. Appl. Environ. Microbiol. 75, 3016–3028 (2009).
Hacker, J. & Kaper, J. B. Pathogenicity islands and the evolution of microbes. Annu. Rev. Microbiol. 54, 641–679 (2000).
Sturino, J. M. & Klaenhammer, T. R. Bacteriophage defense systems and strategies for lactic acid bacteria. Adv. Appl. Microbiol. 56, 331–378 (2004).
Peng, X., Kessler, A., Phan, H., Garrett, R. A. & Prangishvili, D. Multiple variants of the archaeal DNA rudivirus SIRV1 in a single host and a novel mechanism of genomic variation. Mol. Microbiol. 54, 366–375 (2004).
Vestergaard, G. et al. Stygiolobus rod-shaped virus and the interplay of crenarchaeal rudiviruses with the CRISPR antiviral system. J. Bacteriol. 190, 6837–6845 (2008).
Sorek, R., Kunin, V. & Hugenholtz, P. CRISPR — a widespread system that provides acquired resistance against phages in bacteria and archaea. Nature Rev. Microbiol. 6, 181–186 (2008).
Kamerbeek, J. et al. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J. Clin. Microbiol. 35, 907–914 (1997).
Driscoll, J. R. Spoligotyping for molecular epidemiology of the Mycobacterium tuberculosis complex. Methods Mol. Biol. 551, 117–128 (2009).
Mokrousov, I., Limeschenko, E., Vyazovaya, A. & Narvskaya, O. Corynebacterium diphtheriae spoligotyping based on combined use of two CRISPR loci. Biotechnol. J. 2, 901–906 (2007).
Vergnaud, G. et al. Analysis of the three Yersinia pestis CRISPR loci provides new tools for phylogenetic studies and possibly for the investigation of ancient DNA. Adv. Exp. Med. Biol. 603, 327–338 (2007).
Mills, S. et al. CRISPR analysis of bacteriophage-insensitive mutants (BIMs) of industrial Streptococcus thermophilus — implications for starter design. J. Appl. Microbiol.20 Jul 2009 (doi:10.1111/j.1365-2672.2009.04486.x).
Mc Grath, S., Fitzgerald, G. F. & van Sinderen, D. Bacteriophages in dairy products: pros and cons. Biotechnol. J. 2, 450–455 (2007).
Zegans, M. E. et al. Interaction between bacteriophage DMS3 and host CRISPR region inhibits group behaviors of Pseudomonas aeruginosa. J. Bacteriol. 191, 210–219 (2009).
Viswanathan, P., Murphy, K., Julien, B., Garza, A. G. & Kroos, L. Regulation of dev, an operon that includes genes essential for Myxococcus xanthus development and CRISPR-associated genes and repeats. J. Bacteriol. 189, 3738–3750 (2007).
Carthew, R. W. & Sontheimer, E. J. Origins and mechanisms of miRNAs and siRNAs. Cell 136, 642–655 (2009).
Malone, C. D. & Hannon, G. J. Small RNAs as guardians of the genome. Cell 136, 656–668 (2009).
Moazed, D. Small RNAs in transcriptional gene silencing and genome defence. Nature 457, 413–420 (2009).
Siomi, H. & Siomi, M. C. On the road to reading the RNA-interference code. Nature 457, 396–404 (2009).
Han, D., Lehmann, K. & Krauss, G. SSO1450 — a CAS1 protein from Sulfolobus solfataricus P2 with high affinity for RNA and DNA. FEBS Lett. 583, 1928–1932 (2009).
Beloglazova, N. et al. A novel family of sequence-specific endoribonucleases associated with the clustered regularly interspaced short palindromic repeats. J. Biol. Chem. 283, 20361–20371 (2008).
Agari, Y., Yokoyama, S., Kuramitsu, S. & Shinkai, A. X-ray crystal structure of a CRISPR-associated protein, Cse2, from Thermus thermophilus HB8. Proteins 73, 1063–1067 (2008).
Sakamoto, K. et al. X-ray crystal structure of a CRISPR-associated RAMP superfamily protein, Cmr5, from Thermus thermophilus HB8. Proteins 75, 528–532 (2009).
Acknowledgements
We thank R. Terns and M. Terns for communicating results before publication. L.A.M. is a fellow of The Jane Coffin Childs Memorial Fund for Medical Research. This work was supported by a grant from the US National Institutes of Health to E.J.S.
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Glossary
- Transformation
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Genetic alteration of a cell resulting from the acquisition of genes from free DNA molecules in the surrounding environment.
- Conjugation
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The transfer of genetic information from a donor to a recipient cell by a conjugative or mobile genetic element, often a conjugative plasmid.
- Bacteriophage
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(Also abbreviated to 'phage'.) A virus that infects bacteria. Virulent phages kill the host (lytic infection cycle), whereas temperate phages can integrate into the host chromosome (lysogenic cycle), becoming a prophage.
- Transduction
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The transfer of genetic information from one bacterial or archaeal cell to another by a phage particle containing chromosomal DNA.
- DNA restriction
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The destruction of foreign dsDNA by a restriction endonuclease. The protection of self DNA from restriction is achieved by DNA methylation.
- Surface exclusion
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A process that bars conjugative transfer of a plasmid into recipient cells that already harbour a related plasmid.
- RNA interference
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A set of related pathways in eukaryotic cells that use small (20–30 nucleotides) RNAs to regulate the expression or function of cognate sequences.
- Protospacer
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Phage or plasmid sequences that match one or more clustered, regularly interspaced short palindromic repeat (CRISPR) spacer sequences and are targeted during CRISPR interference.
- Dyad symmetry
-
A twofold rotational symmetry relationship (in this case, a DNA arrangement in which a 5′→3′ sequence on one strand is juxtaposed with the same 5′→3′ sequence on the opposite strand). Transcripts from such regions have the capacity to form stem–loop structures.
- Crenarchaeal
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Referring to members of the Crenarchaeota phylum, which is composed mainly of thermophilic archaeal organisms.
- Histidine-aspartate nuclease
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A divalent-metal-dependent phosphohydrolase with a conserved histidine-aspartate motif.
- Unwindase
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Enzyme that uses the free energy of NTP binding and hydrolysis to drive the separation of complementary RNA or DNA strands.
- Virulence factor
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A gene responsible for the production of a molecule that contributes to the establishment of disease by bacterial pathogens.
- Polylysogen
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A lysogen is a bacterium that has a prophage integrated into its chromosome. A polylysogen contains many prophage sequences in its genome.
- Virulence plasmid
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A plasmid that carries virulence factor genes or pathogenicity islands.
- Pathogenicity island
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Genomic islands that contain genes that are required for virulence. These islands are usually absent from non-pathogenic organisms and are acquired by horizontal gene transfer.
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Marraffini, L., Sontheimer, E. CRISPR interference: RNA-directed adaptive immunity in bacteria and archaea. Nat Rev Genet 11, 181–190 (2010). https://doi.org/10.1038/nrg2749
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DOI: https://doi.org/10.1038/nrg2749
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