Abstract:
:The human enteropathogen Clostridium difficile constitutes a key public health issue in industrialized countries. Many aspects of C. difficile pathophysiology and adaptation inside the host remain poorly understood. We have recently reported that this bacterium possesses an active CRISPR-Cas system of subtype I-B for defense against phages and other mobile genetic elements that could contribute to its success during infection. In this paper, we demonstrate that redirecting this endogenous CRISPR-Cas system toward autoimmunity allows efficient genome editing in C. difficile We provide a detailed description of this newly developed approach and show, as a proof of principle, its efficient application for deletion of a specific gene in reference strain 630Δerm and in epidemic C. difficile strain R20291. The new method expands the arsenal of the currently limiting set of gene engineering tools available for investigation of C. difficile and may serve as the basis for new strategies to control C. difficile infections.IMPORTANCEClostridium difficile represents today a real danger for human and animal health. It is the leading cause of diarrhea associated with health care in adults in industrialized countries. The incidence of these infections continues to increase, and this trend is accentuated by the general aging of the population. Many questions about the mechanisms contributing to C. difficile's success inside the host remain unanswered. The set of genetic tools available for this pathogen is limited, and new developments are badly needed. C. difficile has developed efficient defense systems that are directed against foreign DNA and that could contribute to its survival in phage-rich gut communities. We show how one such defense system, named CRISPR-Cas, can be hijacked for C. difficile genome editing. Our results also show a great potential for the use of the CRISPR-Cas system for the development of new therapeutic strategies against C. difficile infections.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Maikova A,Kreis V,Boutserin A,Severinov K,Soutourina Odoi
10.1128/AEM.01416-19subject
Has Abstractpub_date
2019-10-01 00:00:00issue
20eissn
0099-2240issn
1098-5336pii
AEM.01416-19journal_volume
85pub_type
杂志文章abstract::The impact of promoter modification on the expression of the mosquitocidal Bacillus thuringiensis subsp. israelensis cryIVB gene when used to transform the cyanobacterium Synechococcus sp. strain PCC 7942 has been examined. Maximal transcript and protein abundances were achieved by the addition of the lacZ promoter up...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.59.8.2404-2410.1993
更新日期:1993-08-01 00:00:00
abstract::Nutritional and physical factors which influence the germination of akinetes of Nodularia spumigena (Cyanobacteriaceae) were examined. Low concentrations of phosphorus (<0.9 muM) were required for germination. Nitrate had no effect, but ammonia, at concentrations of >45 muM, inhibited germination. Salinities of >20 pe...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.49.1.73-78.1985
更新日期:1985-01-01 00:00:00
abstract::The quorum sensing (QS) system controls bacterial group behaviors in response to cell density. In vibrios, LuxR and AphA are two master QS regulators (MQSRs) controlling gene expression in response to high or low cell density. Other regulators involved in the regulation of these two MQSRs and QS pathways remain to be ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00444-18
更新日期:2018-05-31 00:00:00
abstract::Swimming in ocean water, including ocean water at beaches not impacted by known point sources of pollution, is an increasing health concern. This study was an initial evaluation of the presence of indicator microbes and pathogens and the association among the indicator microbes, pathogens, and environmental conditions...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02127-09
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abstract::Lactobacillus bulgaricus LT4(0448) is a lysogenic strain from which a temperate bacteriophage can be induced by mitomycin C or UV irradiation. Lactobacillus lactis CNRZ 326 is an indicator strain for the temperate phage 0448, but this strain lyses only in the presence of Ca ions. A resistant culture developed secondar...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.53.8.1850-1854.1987
更新日期:1987-08-01 00:00:00
abstract::Algal biofuels represent one of the most promising means of sustainably replacing liquid fuels. However, significant challenges remain before alga-based fuels become competitive with fossil fuels. One of the largest challenges is the ability to harvest the algae in an economical and low-energy manner. In this article,...
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doi:10.1128/AEM.01496-13
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.48.5.994-1000.1984
更新日期:1984-11-01 00:00:00
abstract::Research on the microbial communities that colonize stone monuments may provide a new understanding of stone biodeterioration and microbe-induced carbonate precipitation. This work investigated the seasonal variation of microbial communities in 2016 and 2017, as well as its effects on stone monuments. We determined th...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02680-17
更新日期:2018-03-19 00:00:00
abstract::Actinomycete isolates from indoor air and dust in water-damaged schools and children's day care centers were tested for toxicity by using boar spermatozoa as an indicator. Toxicity was detected in extracts of four strains which caused a loss of sperm motility, and the 50% effective concentrations (EC50) were 10 to 63 ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.64.12.4767-4773.1998
更新日期:1998-12-01 00:00:00
abstract::The ubiquitous arbuscular mycorrhizal fungi consume significant amounts of plant assimilated C, but this C flow has been difficult to quantify. The neutral lipid fatty acid 16:1omega5 is a quantitative signature for most arbuscular mycorrhizal fungi in roots and soil. We measured carbon transfer from four plant specie...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.5.2592-2599.2005
更新日期:2005-05-01 00:00:00
abstract::Hepatitis A virus disinfection was assessed on contaminated stainless-steel disks. Ten microliters of fecally suspended hepatitis A virus was deposited on the center of each disk, dried for 20 min, and then covered with 20 microliters of the test product for 1 min. Of the 20 formulations tested, only 2% glutaraldehyde...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.56.11.3601-3604.1990
更新日期:1990-11-01 00:00:00
abstract::The kinetics of concentration-dependent toluene metabolism were examined by evaluating each term in the second-order rate equation. Marine and freshwater pseudomonads were used. Uptake for Pseudomonas sp. strain T2 was characterized by a completely saturatable system with small transport constant (K(t) = 44 mug/liter)...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.53.9.2193-2205.1987
更新日期:1987-09-01 00:00:00
abstract::Aminopolycarboxylic acids are ubiquitous in natural waters and wastewaters. They have the ability to form very stable water-soluble complexes with many metallic di- or trivalent ions. The iron complex nitrilotriacetic acid-Fe(III) (FeNTA) has been previously shown to increase drastically the rate of photo- and biodegr...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00537-08
更新日期:2008-10-01 00:00:00
abstract::A new design, flat-bottom combination pH electrode was evaluated for utility in determining the pH of prepared agar media by surface contact of the electrode with the agar media. ...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.34.2.242-243.1977
更新日期:1977-08-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.58.1.431-433.1992
更新日期:1992-01-01 00:00:00
abstract::The accumulation and fate of model microbial "pathogens" within a drinking-water distribution system was investigated in naturally grown biofilms formed in a novel pilot-scale water distribution system provided with chlorinated and UV-treated water. Biofilms were exposed to 1-mum hydrophilic and hydrophobic microspher...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.2.706-712.2005
更新日期:2005-02-01 00:00:00
abstract::Nitrate, sulfate, and carbonate were used as electron acceptors to examine the anaerobic biodegradability of chlorinated aromatic compounds in estuarine and freshwater sediments. The respective denitrifying, sulfidogenic, and methanogenic enrichment cultures were established on each of the monochlorinated phenol and m...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.59.4.1162-1167.1993
更新日期:1993-04-01 00:00:00
abstract::Although their exact function remains enigmatic, bifidobacteria are among the first colonizers of the newborn infant gut and further develop into abundant communities, notably in response to diet. Therefore, the transcriptional responses of bifidobacteria in rapidly processed fecal samples from young infants that were...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02492-08
更新日期:2009-05-01 00:00:00
abstract::A gram-negative bacterium isolated from activated sludge was able to utilize up to 25 mM phosphonoacetate as the sole carbon and phosphorus source, with simultaneous excretion of virtually equimolar levels of phosphate. 2-Aminoethylphosphonate was similarly utilized with equivalent growth rates and cellular yields, wh...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.58.4.1364-1366.1992
更新日期:1992-04-01 00:00:00
abstract::Culture-independent (PCR with Crenarchaeota-specific primers and subsequent denaturing gradient gel electrophoresis) and culture-dependent approaches were used to study the diversity of Crenarchaeota in terrestrial hot springs of the Kamchatka Peninsula and the Lake Baikal region (Russia) and of Iceland. Among the phy...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.00972-08
更新日期:2008-12-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.02403-10
更新日期:2011-04-01 00:00:00
abstract::Vibrio cholerae identification based on molecular sequence data has been hampered by a lack of sequence variation from the closely related Vibrio mimicus. The two species share many genes coding for proteins, such as ctxAB, and show almost identical 16S DNA coding for rRNA (rDNA) sequences. Primers targeting conserved...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.65.5.2202-2208.1999
更新日期:1999-05-01 00:00:00
abstract::The diversity of the methanotrophic community in mildly acidic landfill cover soil was assessed by three methods: two culture-independent molecular approaches and a traditional culture-based approach. For the first of the molecular studies, two primer pairs specific for the 16S rRNA gene of validly published type I (i...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.65.11.4887-4897.1999
更新日期:1999-11-01 00:00:00
abstract::Ground bark and heartwood from Alnus rubra and Pseudotsuga menziesii were added to a muddy sediment from a small Oregon stream and incubated in situ. Carbon dioxide and methane production rates were increased by all amendments, the biggest increase being shown with A. rubra wood. Except for sediment amended with A. ru...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.45.2.516-521.1983
更新日期:1983-02-01 00:00:00
abstract::Produce isolates of the Escherichia coli Ont:H52 serotype carried Shiga toxin 1 and stable toxin genes but only expressed Stx1. These strains had pulsed-field gel electrophoresis profiles that were 90% homologous to clinical Ont:H52 strains that had identical phenotypes and genotypes. All Ont:H52 strains had identical...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.72.4.3062-3065.2006
更新日期:2006-04-01 00:00:00
abstract::The cholera toxin genes of Vibrio cholerae are encoded by CTXΦ, a lysogenic bacteriophage. Infection with this phage plays a determinant role in toxigenicity conversion and the emergence of new clones of pathogenic V. cholerae. Multiple phage alleles, defined by sequence types of the repressor gene rstR, have been fou...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.01742-15
更新日期:2015-10-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.66.8.3624-3627.2000
更新日期:2000-08-01 00:00:00
abstract::Nitrite-dependent anaerobic oxidation of methane (n-damo) and ammonium (anammox) are two recently discovered processes in the nitrogen cycle that are catalyzed by n-damo bacteria, including "Candidatus Methylomirabilis oxyfera," and anammox bacteria, respectively. The feasibility of coculturing anammox and n-damo bact...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.05539-11
更新日期:2011-10-01 00:00:00
abstract::The growth of marine bacteria under iron-limited conditions was investigated. Neither siderophore production nor bacterial growth was detected for Pelagiobacter sp. strain V0110 when Fe(III) was present in the culture medium at a concentration of <1.0 microM. However, the growth of V0110 was strongly stimulated by the...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.66.7.2797-2803.2000
更新日期:2000-07-01 00:00:00
abstract::We report here the cloning and sequencing of the gene for proline dehydrogenase (putA) of Bradyrhizobium japonicum. An open reading frame coding for 1,016 amino acids was identified. The B. japonicum gene codes for a bifunctional protein with proline dehydrogenase and pyrroline-5-carboxylate (P5C) dehydrogenase activi...
journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.62.1.221-229.1996
更新日期:1996-01-01 00:00:00