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Non-viral AND transfection

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https://www.readbyqxmd.com/read/27924861/size-specific-transfection-to-mammalian-cells-by-micropillar-array-electroporation
#1
Yingbo Zu, Shuyan Huang, Yang Lu, Xuan Liu, Shengnian Wang
Electroporation serves as a promising non-viral gene delivery approach, while its current configuration carries several drawbacks associated with high-voltage electrical pulses and heterogeneous treatment on individual cells. Here we developed a new micropillar array electroporation (MAE) platform to advance the electroporation-based delivery of DNA and RNA probes into mammalian cells. By introducing well-patterned micropillar array texture on the electrode surface, the number of pillars each cell faces varies with its plasma membrane surface area, despite their large population and random locations...
December 7, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27920474/hepatitis-b-virus-upregulates-host-expression-of-%C3%AE-1-2-mannosidases-via-the-ppar%C3%AE-pathway
#2
Song Hu, Li-Bin Jiang, Xiao-Jing Zou, Wei Yi, De-Ying Tian
AIM: To assess the effects of hepatitis B virus (HBV) on the expression of host α-1,2-mannosidases and determine the underlying mechanisms. METHODS: We measured the expression levels of MAN1A1, MAN1A2, MAN1B1, and MAN1C1 in cell lines HepG2.2.15, HepN10, HepAD38 and HepG2 by Western blot. Viral antigens (HBsAg and HBeAg) in the culture medium were measured using the chemiluminescence method. HBV DNA quantification assays were performed using a commercial real-time PCR kit...
November 21, 2016: World Journal of Gastroenterology: WJG
https://www.readbyqxmd.com/read/27918590/effects-of-circular-dna-length-on-transfection-efficiency-by-electroporation-into-hela-cells
#3
Benjamin D Hornstein, Dany Roman, Lirio M Arévalo-Soliz, Melinda A Engevik, Lynn Zechiedrich
The ability to produce extremely small and circular supercoiled vectors has opened new territory for improving non-viral gene therapy vectors. In this work, we compared transfection of supercoiled DNA vectors ranging from 383 to 4,548 bp, each encoding shRNA against GFP under control of the H1 promoter. We assessed knockdown of GFP by electroporation into HeLa cells. All of our vectors entered cells in comparable numbers when electroporated with equal moles of DNA. Despite similar cell entry, we found length-dependent differences in how efficiently the vectors knocked down GFP...
2016: PloS One
https://www.readbyqxmd.com/read/27903799/cxcr6-mediated-sivagmsab-entry-into-sabaeus-african-green-monkey-lymphocytes-implicates-widespread-use-of-non-ccr5-pathways-in-natural-host-infections
#4
Katherine S Wetzel, Yanjie Yi, Sarah T C Elliott, Dino Romero, Beatrice Jacquelin, Beatrice H Hahn, Michaela Muller-Trutwin, Cristian Apetrei, Ivona Pandrea, Ronald G Collman
: African green monkeys (AGM) and sooty mangabeys (SM) are well-studied natural hosts of SIV, which do not progress to AIDS when infected with their species-specific viruses. SIV natural hosts express very low levels of the canonical entry coreceptor CCR5, and recent studies have shown that CCR5 is dispensable for SIV infection of SM in vivo, and blocking CCR5 does not prevent ex vivo infection of PBMC from SM or vervet AGM. In both hosts, CXCR6 is an efficient entry pathway in vitro...
November 30, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27900567/clinical-potential-of-human-induced-pluripotent-stem-cells-perspectives-of-induced-pluripotent-stem-cells
#5
REVIEW
Dharmendra Kumar, Taruna Anand, Wilfried A Kues
The recent establishment of induced pluripotent stem (iPS) cells promises the development of autologous cell therapies for degenerative diseases, without the ethical concerns associated with human embryonic stem (ES) cells. Initially, iPS cells were generated by retroviral transduction of somatic cells with core reprogramming genes. To avoid potential genotoxic effects associated with retroviral transfection, more recently, alternative non-viral gene transfer approaches were developed. Before a potential clinical application of iPS cell-derived therapies can be planned, it must be ensured that the reprogramming to pluripotency is not associated with genome mutagenesis or epigenetic aberrations...
November 29, 2016: Cell Biology and Toxicology
https://www.readbyqxmd.com/read/27899629/enhanced-non-viral-gene-delivery-by-coordinated-endosomal-release-and-inhibition-of-%C3%AE-tubulin-deactylase
#6
Yoon Khei Ho, Li Han Zhou, Kam C Tam, Heng Phon Too
Efficient non-viral gene delivery is highly desirable but often unattainable with some cell-types. We report here that non-viral DNA polyplexes can efficiently transfect differentiated neuronal and stem cells. Polyplex transfection centrifugation protocols was enhanced by including a simultaneous treatment with a DOPE/CHEMS lipid suspension and a microtubule inhibitor, Tubastatin A. Lipoplex transfection protocols were not improved by this treatment. This mechanism of action was unravelled by systematically identifying and rationally mitigating barriers limiting high transfection efficiency, allowing unexpected improvements in the transfection of mesenchymal stem cells (MSC), primary neuron and several hard-to-transfect cell types beyond what are currently achievable using cationic polymers...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27885808/vp1-the-major-capsid-protein-of-the-mouse-polyomavirus-binds-microtubules-promotes-their-acetylation-and-blocks-the-host-cell-cycle
#7
Lenka Horníková, Martin Fraiberk, Petr Man, Václav Janovec, Jitka Forstová
VP1, the major structural protein of the mouse polyomavirus (MPyV), is the major architectural component of the viral capsid. Its pentamers are able to self-assemble into capsid-like particles and to non-specifically bind DNA. Surface loops of the protein interact with sialic acid of ganglioside receptors. Although the replication cycle of the virus, including virion morphogenesis, proceeds in the cell nucleus, a substantial fraction of the protein is detected in the cytoplasm of late-phase MPyV-infected cells...
November 25, 2016: FEBS Journal
https://www.readbyqxmd.com/read/27878462/enterovirus-a71-and-coxsackievirus-a16-show-different-replication-kinetics-in-human-neuronal-and-non-neuronal-cell-lines
#8
Thinesshwary Yogarajah, Kien Chai Ong, David Perera, Kum Thong Wong
Enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16) are closely related enteroviruses that cause hand, foot and mouth disease (HFMD) in children. Serious neurological complications almost always occur in EV-A71 infection, but are rare in CV-A16 infection. Based on the hypothesis that this may be because EV-A71 infects neuronal cells more easily than CV-A16, we compared virus infection, replication and spread of EV-A71 and CV-A16 in SK-N-SH cells. We found that CV-A16 invariably showed significantly lower replication and caused less necrotic cell death in SK-N-SH cells, compared with EV-A71...
November 23, 2016: Archives of Virology
https://www.readbyqxmd.com/read/27873273/isolation-and-characterization-of-exosomes-released-by-ebv-immortalized-cells
#9
Gulfaraz Khan, Waqar Ahmed
Epstein-Barr virus is an oncogenic herpesvirus associated with several human malignancies. Although the details of the molecular steps involved in EBV-mediated cell transformation and immune evasion are not fully known, a number of viral products, including EBV latent proteins and non-protein coding RNAs have been shown to be involved, directly or indirectly in these processes. In recent years, a growing body of data indicates that some viruses are able to transport selected products to neighboring cells and induce biological changes by exploiting the exosome secretory pathway...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27872706/alkyl-cross-linked-low-molecular-weight-polypropyleneimine-dendrimers-as-efficient-gene-delivery-vectors
#10
Faezeh Moghadam Ariaee, Maryam Hashemi, Sara Amel Farzad, Khalil Abnous, Mohammad Ramezani
OBJECTIVES: In recent years, polypropyleneimine (PPI) dendrimers have attracted great interest as non-viral gene delivery systems because of their attractive features including highly branched architecture with number of reactive end groups. However, without being structurally modified, they are not efficient gene carriers. In the present study, generation 2 and 3 (G2 and G3) of PPI dendrimers were conjugated with alkylcarboxylate groups as linker to enhance the transfection efficiency while maintaining their low cell toxicity...
October 2016: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/27872619/ebola-vp40-in-exosomes-can-cause-immune-cell-dysfunction
#11
Michelle L Pleet, Allison Mathiesen, Catherine DeMarino, Yao A Akpamagbo, Robert A Barclay, Angela Schwab, Sergey Iordanskiy, Gavin C Sampey, Benjamin Lepene, Sergei Nekhai, M J Aman, Fatah Kashanchi
Ebola virus (EBOV) is an enveloped, ssRNA virus from the family Filoviridae capable of causing severe hemorrhagic fever with up to 80-90% mortality rates. The most recent outbreak of EBOV in West Africa starting in 2014 resulted in over 11,300 deaths; however, long-lasting persistence and recurrence in survivors has been documented, potentially leading to further transmission of the virus. We have previously shown that exosomes from cells infected with HIV-1, HTLV-1 and Rift Valley Fever virus are able to transfer viral proteins and non-coding RNAs to naïve recipient cells, resulting in an altered cellular activity...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27871835/development-of-flexible-nanocarriers-for-sirna-delivery-into-tumor-tissue
#12
Hyunkyung Jung, Yuri Shimatani, Mahadi Hasan, Kohei Uno, Susumu Hama, Kentaro Kogure
Various non-viral delivery systems for small interfering RNAs (siRNA) have been developed. Such delivery systems generally exhibit tightly formed spherical structures. While such carriers have demonstrated good transfection activity in mono-layered cell systems, effects against solid tumors are often less apparent and difficult to demonstrate, likely due to the rigid structures of the carriers, which may prevent penetration to deeper regions within tumor tissue. Herein, we developed a flexible nanocarrier (FNC) system that is able to penetrate to deeper regions within tumor tissue...
November 18, 2016: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/27868311/tnf-%C3%AE-induced-il-6-and-mmp-9-expression-in-immortalized-ameloblastoma-cell-line-established-by-htert
#13
K Ohta, T Naruse, Y Ishida, H Shigeishi, T Nakagawa, A Fukui, H Nishi, K Sasaki, I Ogawa, M Takechi
OBJECTIVE: Ameloblastoma (AM) shows locally invasive behaviour. However, biological investigations regarding regulation of gene expression associated with AM pathological features are difficult to perform, because AM cells can be passaged for a few generations due to senescence. We report a newly established immortalized AM cell line, AMB cells, by transfection with human telomerase reverse transcriptase (hTERT). Furthermore, we examined whether TNF-α modulates bone resorption-related genes, IL-6 and MMP-9 in cooperation with TGF-β or IFN-γ...
October 13, 2016: Oral Diseases
https://www.readbyqxmd.com/read/27867077/critical-phases-of-viral-production-processes-monitored-by-capacitance
#14
E Petiot, S Ansorge, Manuel Rosa-Calatrava, A Kamen
Over the last decade industrial manufacturing of viral vaccines and viral vectors for prophylactic and therapeutic applications is experiencing a remarkable growth. Currently, the quality attributes of viral derived products are assessed only at the end-point of the production process, essentially because in-process monitoring tools are not available or not implemented at industrial scale. However, to demonstrate process reproducibility and robustness, manufacturers are strongly advised by regulatory agencies to adopt more on-line process monitoring and control...
November 17, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27866318/pig-bvdv-2-non-structural-protein-n-pro-links-to-cellular-antiviral-response-in-vitro
#15
Jie Tao, Jinhu Liao, Jianye Wang, Xinjun Zhang, Qian Zhang, Liqian Zhu, Weiping Zhang, Huili Liu, Guoqiang Zhu
In this study, we constructed for the first time a full-length cDNA clone of pig-original bovine viral diarrhea virus 2 (BVDV-2) strain SH-28, modified the cDNA clone (pASH28) for mutant pASHΔN(pro) and derived virus strain vASHΔN(pro) by deleting the genomic region encoding the N(pro) polypeptide, and examined significance of protein N(pro) for antiviral responses in vitro. Data showed that N(pro)-deletion mutant virus vASHΔN(pro) led to significant overexpression of oligo adenylate synthetase (OAS), myxovirus-resistant protein 1 (Mx1), and ubiquitin-like protein 15 (ISG15)...
November 19, 2016: Virus Genes
https://www.readbyqxmd.com/read/27860328/luminescent-ruthenium-ii-polypyridyl-complexes-as-non-viral-carriers-for-dna-delivery
#16
Satish Shantaram Bhat, Vidyanand K Revankar, Ayesha Khan, Rahul V Pinjari, Marek Necas
Two new luminescent ruthenium(II) polypyridyl complexes [Ru(bpy)2(tpt-phen)]Cl2 (1) and [Ru(phen)2(tpt-phen)]Cl2 (2) {bpy = 2,2'-bipyridine, phen = 1,10-phenanthroline, tpt-phen = triptycenyl-1,10-phenanthroline} have been developed as potential non-viral vectors for DNA delivery. Photophysical and electrochemical properties of the complexes have been investigated and corroborated with electronic structure calculations. The DNA condensation by these complexes has been investigated by UV-visible and emission spectroscopy, circular dichroism spectroscopy, atomic force microscopy, dynamic light scattering, confocal microscopy and electrophoretic mobility studies...
November 10, 2016: Chemistry, An Asian Journal
https://www.readbyqxmd.com/read/27856695/multiplex-shrna-screening-of-germ-cell-development-by-in-vivo-transfection-of-mouse-testis
#17
Nicholas R Y Ho, Abul R Usmani, Yan Yin, Liang Ma, Donald F Conrad
Spermatozoa are one of the few mammalian cells types that cannot be fully derived in vitro, severely limiting the application of modern genomic techniques to study germ cell biology. The current gold standard approach of characterizing single gene knockout mice is slow as generation of each mutant line can take 6-9 months. Here, we describe an in vivo approach to rapid functional screening of germline genes based on a new non-surgical, non-viral in vivo transfection method to deliver nucleic acids into testicular germ cells...
November 16, 2016: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/27837544/preparation-and-physical-characterization-of-dna-binding-cationic-liposomes
#18
Vaibhav Saxena
DNA-binding cationic liposomes are routinely employed as one of the major non-viral transfecting agents delivering DNA and other genes inside the cells. Cationic liposomes when complexed with DNA form a strong positively charged liposome-DNA complex or lipoplex. The chapter discusses primarily the major preparation technique for cationic liposomes and its physical characterization, with a focus on SYBR Green-I dye exclusion assay and DNA encapsulation enhancement by freeze-thaw technique. SYBR Green-I dye exclusion assay is a technique to determine the total amount of liposomal lipids required to bind a unit weight of DNA, which is critical for transfection experiments...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27826075/the-effect-of-the-dengue-non-structural-1-protein-expression-over-the-hepg2-cell-proteins-in-a-proteomic-approach
#19
Kíssila Rabelo, Monique R O Trugilho, Simone M Costa, Bernardo A S Pereira, Otacílio C Moreira, André T S Ferreira, Paulo C Carvalho, Jonas Perales, Ada M B Alves
: Dengue is an important mosquito borne viral disease in the world. Dengue virus (DENV) encodes a polyprotein, which is cleaved in ten proteins, including the non-structural protein 1 (NS1). In this work, we analyzed the effect of NS1 expression in one hepatic cell line, HepG2, through a shotgun proteomic approach. Cells were transfected with pcENS1 plasmid, which encodes the DENV2 NS1 protein, or the controls pcDNA3 (negative control) and pMAXGFP (GFP, a protein unrelated to dengue)...
November 5, 2016: Journal of Proteomics
https://www.readbyqxmd.com/read/27824330/effect-of-sustained-pdgf-non-viral-gene-delivery-on-repair-of-tooth-supporting-bone-defects
#20
A B Plonka, B Khorsand, N Yu, J V Sugai, A K Salem, W V Giannobile, S Elangovan
Recombinant human platelet-derived growth factor-BB (rhPDGF-BB) promotes soft tissue and bone healing, and is FDA-approved for treatment of diabetic ulcers and periodontal defects. The short half-life of topical rhPDGF-BB protein application necessitates bolus, high-dose delivery. Gene therapy enables sustained local growth factor production. A novel gene activated matrix delivering polyplexes of polyethylenimine (PEI)-plasmid DNA (pDNA) encoding PDGF was evaluated for promotion of periodontal wound repair in vivo...
November 8, 2016: Gene Therapy
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