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Plasmid curing

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https://www.readbyqxmd.com/read/29242561/hepatitis-b-virus-persistence-in-mice-reveals-il-21-and-il-33-as-regulators-of-viral-clearance
#1
Zhongliang Shen, Huijuan Yang, Sisi Yang, Wei Wang, Xiaoxian Cui, Xian Zhou, Wei Liu, Shaokun Pan, Yanfeng Liu, Junqi Zhang, Jiming Zhang, Youhua Xie, Jing Liu
Hepatitis B virus (HBV) generally causes self-limiting infection in immunocompetent adults, but establishes chronic infection in some adults and in most maternally infected infants. Factors determining clearance versus persistence are not fully understood. Hydrodynamic injection (HDI) of HBV replicon plasmid via tail vein generally results in quick clearance in immunocompetent adult mice. Here, we report the identification of strain-specific persistence of HBV in mice: one genotype B strain, designated BPS, persisted up to 33 weeks in ~50% of HDI mice...
December 14, 2017: Nature Communications
https://www.readbyqxmd.com/read/29183134/-b-i-in-vitro-i-transduction-and-target-mutagenesis-efficiency-of-hiv-1-i-pol-i-gene-targeting-zfn-and-crisper-cas9-delivered-by-various-plasmids-and-or-vectors-towards-an-hiv-cure-b
#2
Moses Okee, Alice Bayiyana, Carol Musubika, Moses Joloba, Fred Ashaba-Katabazi, Bernard Ssentalo Bagaya, Misaki Wayengera
Efficiency of artificial restriction enzymes (AREs) towards curing HIV has only been separately examined, using differing delivery vehicles. We compared the in-vitro transduction and target-mutagenesis efficiency of consortium-plasmid and adenoviral vector delivered HIV-1<i> pol </i>gene targeting ZFN with CRISPER/Cas. Custom-ZFN, CRISPER-Cas-9, plasmids and vectors (murCTSD_pZFN, pGS-U-gRNA, pCMV-Cas-D01A, Ad5-RGD); cell-lines (TZM-bl and ACH-2/J-Lat cells); and the latency reversing agents Prostatin, SAHA, and PMA were used...
November 28, 2017: AIDS Research and Human Retroviruses
https://www.readbyqxmd.com/read/29162712/persistence-of-an-oncogenic-papillomavirus-genome-requires-cis-elements-from-the-viral-transcriptional-enhancer
#3
Koenraad Van Doorslaer, Dan Chen, Sandra Chapman, Jameela Khan, Alison A McBride
Human papillomavirus (HPV) genomes are replicated and maintained as extrachromosomal plasmids during persistent infection. The viral E2 proteins are thought to promote stable maintenance replication by tethering the viral DNA to host chromatin. However, this has been very difficult to prove genetically, as the E2 protein is involved in transcriptional regulation and initiation of replication, as well as its assumed role in genome maintenance. This makes mutational analysis of viral trans factors and cis elements in the background of the viral genome problematic and difficult to interpret...
November 21, 2017: MBio
https://www.readbyqxmd.com/read/29157810/camptothecin-producing-endophytic-bacteria-from-pyrenacantha-volubilis-hook-icacinaceae-a-possible-role-of-a-plasmid-in-the-production-of-camptothecin
#4
K N Soujanya, R Siva, P Mohana Kumara, Amitava Srimany, G Ravikanth, F A Mulani, T Aarthy, H V Thulasiram, T R Santhoshkumar, Karaba N Nataraja, R Uma Shaanker
BACKGROUND: Camptothecin (CPT), a quinoline alkaloid, is a potent inhibitor of eukaryotic topoisomerase I. Because of this property, several derivatives of CPT are used as chemotherapeutic agents. CPT is produced by several plant species belonging to the Asterid clade as well as by a number of endophytic fungal associates of these plants. In this study, we report the production of CPT by four bacterial endophytes and show the possible role of a plasmid in the biosynthesis of CPT. METHODS: Endophytic bacteria were isolated from leaves, stems and fruits of Pyrenacantha volubilis Hook...
December 1, 2017: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
https://www.readbyqxmd.com/read/29134663/multiple-plasmid-borne-virulence-genes-of-clavibacter-michiganensis-ssp-capsici-critical-for-disease-development-in-pepper
#5
In Sun Hwang, Eom-Ji Oh, Donghyuk Kim, Chang-Sik Oh
Clavibacter michiganensis ssp. capsici is a Gram-positive plant-pathogenic bacterium causing bacterial canker disease in pepper. Virulence genes and mechanisms of C. michiganensis ssp. capsici in pepper have not yet been studied. To identify virulence genes of C. michiganensis ssp. capsici, comparative genome analyses with C. michiganensis ssp. capsici and its related C. michiganensis subspecies, and functional analysis of its putative virulence genes during infection were performed. The C. michiganensis ssp...
November 14, 2017: New Phytologist
https://www.readbyqxmd.com/read/29104933/heat-resistance-mediated-by-plm58-plasmid-borne-clpl-in-listeria-monocytogenes
#6
Anna Pöntinen, Mariella Aalto-Araneda, Miia Lindström, Hannu Korkeala
Listeria monocytogenes is one of the most heat-resistant non-spore-forming food-borne pathogens and poses a notable risk to food safety, particularly when mild heat treatments are used in food processing and preparation. While general heat stress properties and response mechanisms of L. monocytogenes have been described, accessory mechanisms providing particular L. monocytogenes strains with the advantage of enhanced heat resistance are unknown. Here, we report plasmid-mediated heat resistance of L. monocytogenes for the first time...
November 2017: MSphere
https://www.readbyqxmd.com/read/29042928/long-non-coding-rna-ab209630-suppresses-cell-proliferation-and-metastasis-in-human-hepatocellular-carcinoma
#7
Teng Li, Yun Liu, Yanming Sun
Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide and the second most common cause of cancer-related mortalities. With a high potential for metastasis and recurrence, HCC is refractory to cure. The present study aimed to explore the role of a recent-discovered LncRNA, AB209630, in human HCC, in order to provide new insights useful for clinical HCC diagnosis and treatment. Reverse transcription-quantitative polymerase chain reaction was performed to examine the expression of AB209630 in clinical HCC samples and the adjacent non-cancerous tissues...
October 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/29027367/the-parasporal-crystals-of-bacillus-pumilus-strain-15-1-a-potential-virulence-factor
#8
Diana C Garcia-Ramon, Colin Berry, Carmen Tse, Alberto Fernández-Fernández, Antonio Osuna, Susana Vílchez
Bacillus pumilus strain 15.1 was previously found to cause larval mortality in the Med-fly Ceratitis capitata and was shown to produce crystals in association with the spore. As parasporal crystals are well-known as invertebrate-active toxins in entomopathogenic bacteria such as Bacillus thuringiensis (Cry and Cyt toxins) and Lysinibacillus sphaericus (Bin and Cry toxins), the B. pumilus crystals were characterized. The crystals were composed of a 45 kDa protein that was identified as an oxalate decarboxylase by peptide mass fingerprinting, N-terminal sequencing and by comparison with the genome sequence of strain 15...
October 12, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/29019352/hot-news-gene-therapy-with-crispr-cas9-coming-to-age-for-hiv-cure
#9
Vicente Soriano
The huge success of current antiretroviral therapy is mediated by a triple effect: (i) Halting progression to AIDS in infected persons; (ii) reducing the risk of transmission to contacts (treatment as prevention); and (iii) minimizing the risk of HIV acquisition treating uninfected persons at risk (pre-exposure prophylaxis). However, UNAIDS has estimated that only 70% of infected people globally are diagnosed, only 53% are treated, and overall 44% have undetectable viral load, which is the necessary request for ensuring any antiretroviral benefit...
October 2017: AIDS Reviews
https://www.readbyqxmd.com/read/29018834/in-situ-liver-expression-of-hbsag-cd3-bispecific-antibodies-for-hbv-immunotherapy
#10
Robert L Kruse, Thomas Shum, Xavier Legras, Mercedes Barzi, Frank P Pankowicz, Stephen Gottschalk, Karl-Dimiter Bissig
Current therapies against hepatitis B virus (HBV) do not reliably cure chronic infection, necessitating new therapeutic approaches. The T cell response can clear HBV during acute infection, and the adoptive transfer of antiviral T cells during bone marrow transplantation can cure patients of chronic HBV infection. To redirect T cells to HBV-infected hepatocytes, we delivered plasmids encoding bispecific antibodies directed against the viral surface antigen (HBsAg) and CD3, expressed on almost all T cells, directly into the liver using hydrodynamic tail vein injection...
December 15, 2017: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/28922955/comparison-of-zinc-finger-nucleases-versus-crispr-specific-nucleases-for-genome-editing-of-the-wiskott-aldrich-syndrome-locus
#11
Alejandra Gutierrez-Guerrero, Sabina Sanchez-Hernandez, Giuseppe Galvani, Javier Pinedo-Gomez, Rocio Martin-Guerra, Almudena Sanchez-Gilabert, Araceli Aguilar-González, Marién Cobo, Philip Gregory, Michael Holmes, Karim Benabdellah, Francisco Martin
Primary immunodeficiencies, including Wiskott-Aldrich syndrome (WAS), are a main target for genome-editing strategies using specific nucleases (SNs) because a small number of corrected hematopoietic stem cells could cure patients. In this work, we have designed various WAS gene-specific CRISPR/Cas9 systems and compared their efficiency and specificity with homodimeric and heterodimeric WAS-specific zinc finger nucleases (ZFNs), using K-562 cells as a cellular model and plasmid nucleofection or integration-deficient lentiviral vectors (IDLVs) for delivery...
October 25, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28918634/engineered-probiotic-for-the-inhibition-of-salmonella-via-tetrathionate-induced-production-of-microcin-h47
#12
Jacob D Palmer, Emma Piattelli, Beth A McCormick, Mark W Silby, Christopher J Brigham, Vanni Bucci
Complications arising from antibiotic-resistant bacteria are becoming one of the key issues in modern medicine. Members of drug-resistant Enterobacteriaceae spp. include opportunistic pathogens (e.g., Salmonella spp.) that are among the leading causes of morbidity and mortality worldwide. Overgrowth of these bacteria is considered a hallmark of intestinal dysbiosis. Microcins (small antimicrobial peptides) produced by some gut commensals can potentially cure these conditions by inhibiting these pathogens and have been proposed as a viable alternative to antibiotic treatment...
September 20, 2017: ACS Infectious Diseases
https://www.readbyqxmd.com/read/28888103/using-crispr-cas-systems-as-antimicrobials
#13
REVIEW
David Bikard, Rodolphe Barrangou
Although CRISPR-Cas systems naturally evolved to provide adaptive immunity in bacteria and archaea, Cas nucleases can be co-opted to target chromosomal sequences rather than invasive genetic elements. Although genome editing is the primary outcome of self-targeting using CRISPR-based technologies in eukaryotes, self-targeting by CRISPR is typically lethal in bacteria. Here, we discuss how DNA damage introduced by Cas nucleases in bacteria can efficiently and specifically lead to plasmid curing or drive cell death...
September 5, 2017: Current Opinion in Microbiology
https://www.readbyqxmd.com/read/28856222/comparison-of-clinical-methods-for-detecting-carbapenem-resistant-enterobacteriaceae
#14
Kirsten Kost, Jumi Yi, Beverly Rogers, Robert Jerris
We evaluated detection of carbapenem-resistant Enterobacteriaceae (CRE) by routine minimal inhibitory concentration (MIC) testing, polymerase chain reaction (PCR) using Xpert® Carba-R assay, hydrolysis of ertapenem and imipenem detected by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS), and hydrolysis by colorimetry using the EPI-CRE assay. Ninety-six Enterobacteriaceae isolates possessing carbapenemase genes and 29 carbapenem-susceptible Enterobacteriaceae were available for testing...
August 2017: Practical Laboratory Medicine
https://www.readbyqxmd.com/read/28854680/the-ctx-m-14-plasmid-phk01-encodes-novel-small-rnas-and-influences-host-growth-and-motility
#15
Xinlei Jiang, Xuan Liu, Carmen O K Law, Ya Wang, Wai U Lo, Xing Weng, Ting Fung Chan, P L Ho, Terrence C K Lau
The dissemination of extended-spectrum β-lactamases (ESBLs) genes among bacteria is commonly achieved by plasmid conjugation. In the last decade, the CTX-M type enzyme was the most widespread and prevalent ESBLs in the world. In Hong Kong and mainland China, among the commonly found CTX-M-carrying plasmids were pHK01 and pHK01-like plasmids, which belong to incompatibility group FII (IncFII). In this work, we studied the physiological effect caused by the pHK01 plasmid in bacterial host Escherichia coli J53...
July 1, 2017: FEMS Microbiology Ecology
https://www.readbyqxmd.com/read/28842543/lactolisterin-bu-a-novel-class-ii-broad-spectrum-bacteriocin-from-lactococcus-lactis-subsp-lactis-bv-diacetylactis-bgbu1-4
#16
Jelena Lozo, Nemanja Mirkovic, Paula M O'Connor, Milka Malesevic, Marija Miljkovic, Natalija Polovic, Branko Jovcic, Paul D Cotter, Milan Kojic
Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 produces a novel bacteriocin, lactolisterin BU, with strong antimicrobial activity against many species of Gram-positive bacteria, including important food spoilage and foodborne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, Bacillus spp., and streptococci. Lactolisterin BU was extracted from the cell surface of BGBU1-4 by 2-propanol and purified to homogeneity by C18 solid-phase extraction and reversed-phase high-performance liquid chromatography...
November 1, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28771369/isolation-and-characterization-of-2-4-d-butyl-ester-degrading-acinetobacter-sp-zx02-from-a-chinese-ginger-cultivated-soil
#17
Lin Xiao, Hai-Fei Jia, In-Hong Jeong, Young-Joon Ahn, Yong-Zhe Zhu
Strain ZX02 was isolated from Chinese ginger cultivated soil contaminated with various pesticides, which could utilize 2,4-dichlorophenoxyacetic acid butyl ester (2,4-D butyl ester) as the sole carbon source. On the basis of the sequence analysis of 16S rRNA gene as well as the morphological, biochemical, and physiological characteristics of strain ZX02, the organism belonged to Gram-negative bacterium and was identified as Acinetobacter sp. ZX02. The strain ZX02 showed a remarkable performance in 2,4-D butyl ester degradation (100% removal in <96 h) in pure culture...
August 30, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28771154/multiresistant-opportunistic-pathogenic-bacteria-isolated-from-polluted-rivers-and-first-detection-of-nontuberculous-mycobacteria-in-the-algerian-aquatic-environment
#18
Lydia Neïla Djouadi, Okba Selama, Ahmed Abderrahmani, Amel Bouanane-Darenfed, Lamia Abdellaziz, Meriam Amziane, Marie-Laure Fardeau, Farida Nateche
Opportunistic infections constitute a major challenge for modern medicine mainly because the involved bacteria are usually multiresistant to antibiotics. Most of these bacteria possess remarkable ability to adapt to various ecosystems, including those exposed to anthropogenic activities. This study isolated and identified 21 multiresistant opportunistic bacteria from two polluted rivers, located in Algiers. Cadmium, lead, and copper concentrations were determined for both water samples to evaluate heavy metal pollution...
August 2017: Journal of Water and Health
https://www.readbyqxmd.com/read/28764701/a-versatile-one-step-crispr-cas9-based-approach-to-plasmid-curing
#19
Ida Lauritsen, Andreas Porse, Morten O A Sommer, Morten H H Nørholm
BACKGROUND: Plasmids are widely used and essential tools in molecular biology. However, plasmids often impose a metabolic burden and are only temporarily useful for genetic engineering, bio-sensing and characterization purposes. While numerous techniques for genetic manipulation exist, a universal tool enabling rapid removal of plasmids from bacterial cells is lacking. RESULTS: Based on replicon abundance and sequence conservation analysis, we show that the vast majority of bacterial cloning and expression vectors share sequence similarities that allow for broad CRISPR-Cas9 targeting...
August 2, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28690290/rescue-of-an-intracellular-avirulent-rhodococcus-equi-replication-defect-by-the-extracellular-addition-of-virulence-associated-protein-a
#20
Nuttapone Sangkanjanavanich, Masanori Kawai, Tsutomu Kakuda, Shinji Takai
Rhodococcus equi is a facultative intracellular bacterium that can escape from bactericidal mechanisms associated with phagocytosis. Virulence-associated protein A (VapA), encoded on a virulence-associated plasmid, is essential for intracellular survival in macrophages, but its function is not known. Here, we show that the extracellular addition of recombinant glutathione S-transferase (GST)-VapA fusion protein rescued the intracellular replication defect of a mutant lacking the vapA gene. Furthermore, the virulence-plasmid-cured strain could also multiply to nearly wild-type levels by the addition of GST-VapA...
August 4, 2017: Journal of Veterinary Medical Science
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