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https://www.readbyqxmd.com/read/28453254/gene-transfection-mediated-by-catiomers-requires-free-highly-charged-polymer-chains-to-overcome-intracellular-barriers
#1
Lindomar Jose Calumby Albuquerque, Carlos E de Castro, Karin A Riske, Maria Cristina Carlan da Silva, Paulo I R Muraro, Vanessa Schmidt Giacomelli, Cristiano Giacomelli, Fernando Carlos Carlos Giacomelli
The prospective use of the block copolymers PEO113-b-PDEA50 and POEGMA70-b-P(OEGMA10-co-DEA47-co-DPA47) as non-viral gene vectors was evaluated. The polymers are able to properly condense DNA into nano-sized particles (RH ~ 75 nm) which are marginally cytotoxic and can be uptaken by cells. However, the GFP expression assays evidenced that DNA delivery is essentially negligible meaning that intracellular trafficking hampers efficient gene release. Subsequently we demonstrate that cellular uptake and particularly the quantity of GFP-positive cells are substantially enhanced when the block copolymer polyplexes are produced and further supplemented by BPEI chains (branched polyethyleneimine)...
April 28, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28447465/transferrin-dressed-virus-like-ternary-nanoparticles-with-aggregation-induced-emission-for-targeted-delivery-and-rapid-cytosolic-release-of-sirna
#2
Tingbin Zhang, Weisheng Guo, Chunqiu Zhang, Jing Yu, Jing Xu, Shuyi Li, Jian-Hua Tian, Paul C Wang, Jinfeng Xing, Xing-Jie Liang
Viruses have evolved to be outstandingly efficient at gene delivery, but their use as vectors is limited by safety risks. Inspired by the structure of viruses, we constructed a virus-mimicking vector (denoted as TR4@siRNA@Tf NCs) with virus-like architecture and infection properties. Composed of a hydrophilic peptide, an AIEgen and a lipophilic tail, TR4 imitates the viral capsid and endows the vector with aggregation-induced emission (AIE) properties as well as efficient siRNA compaction. The outer glycoprotein transferrin (Tf) mimics the viral envelope protein, and endows the vector with reduced cytotoxicity as well as enhanced targeting capability...
April 27, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28443609/the-hiv-1-viral-synapse-signals-human-foreskin-keratinocytes-to-secrete-thymic-stromal-lymphopoietin-facilitating-hiv-1-foreskin-entry
#3
Z Zhou, L Xu, A Sennepin, C Federici, Y Ganor, D Tudor, D Damotte, N Barry Delongchamps, M Zerbib, M Bomsel
The complexity of signal transduction resulting from the contact of human immunodeficiency virus type 1 (HIV-1)-infected cells and mucosal cells has hampered our comprehension of HIV-1 mucosal entry. Such process is driven efficiently only by viral synapse contacts, whereas cell-free HIV-1 remains poorly infectious. Using CD4(+) T-cells expressing only HIV-1 envelope inoculated on human adult foreskin tissues, we designed methodologies to identify the signals transduced in foreskin keratinocytes following HIV-1-envelope-dependent viral synapse formation...
April 26, 2017: Mucosal Immunology
https://www.readbyqxmd.com/read/28442771/double-stranded-dna-induces-a-prothrombotic-phenotype-in-the-vascular-endothelium
#4
Erik Gaitzsch, Thomas Czermak, Andrea Ribeiro, Yvonn Heun, Monica Bohmer, Monika Merkle, Hanna Mannell, Christian Schulz, Markus Wörnle, Joachim Pircher
Double-stranded DNA (dsDNA) constitutes a potent activator of innate immunity, given its ability to bind intracellular pattern recognition receptors during viral infections or sterile tissue damage. While effects of dsDNA in immune cells have been extensively studied, dsDNA signalling and its pathophysiological implications in non-immune cells, such as the vascular endothelium, remain poorly understood. The aim of this study was to characterize prothrombotic effects of dsDNA in vascular endothelial cells. Transfection of cultured human endothelial cells with the synthetic dsDNA poly(dA:dT) induced upregulation of the prothrombotic molecules tissue factor and PAI-1, resulting in accelerated blood clotting in vitro, which was partly dependent on RIG-I signalling...
April 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28437468/role-of-cleavage-at-the-core-e1-junction-of-hepatitis-c-virus-polyprotein-in-viral-morphogenesis
#5
Véronique Pène, Matthieu Lemasson, Francis Harper, Gérard Pierron, Arielle R Rosenberg
In hepatitis C virus (HCV) polyprotein sequence, core protein terminates with E1 envelope signal peptide. Cleavage by signal peptidase (SP) separates E1 from the complete form of core protein, anchored in the endoplasmic reticulum (ER) membrane by the signal peptide. Subsequent cleavage of the signal peptide by signal-peptide peptidase (SPP) releases the mature form of core protein, which preferentially relocates to lipid droplets. Both of these cleavages are required for the HCV infectious cycle, supporting the idea that HCV assembly begins at the surface of lipid droplets, yet SPP-catalyzed cleavage is dispensable for initiation of budding in the ER...
2017: PloS One
https://www.readbyqxmd.com/read/28433538/the-ns1-protein-from-influenza-virus-stimulates-translation-initiation-by-enhancing-ribosome-recruitment-to-mrnas
#6
Baptiste Panthu, Olivier Terrier, Coralie Carron, Aurélien Traversier, Antoine Corbin, Laurent Balvay, Bruno Lina, Manuel Rosa-Calatrava, Théophile Ohlmann
The non-structural protein NS1 of influenza A viruses exerts pleiotropic functions during infection. Amongst these functions, NS1 was shown to be involved both in the control of viral and cellular translation; however the mechanism by which this occurs remains to be determined. Thus, we have revisited the role of NS1 in translation by using a combination of influenza infection, mRNA reporter transfection, in vitro functional and biochemical assays. Our data show that the NS1 protein is able to enhance translation of virtually all tested mRNAs with the exception of constructs bearing the Dicistroviruses IRESes (DCV and CrPV), suggesting a role at the level of translation initiation...
April 19, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28426973/non-linear-enhancement-of-mrna-delivery-efficiencies-by-influenza-a-derived-ns1-protein-engendering-host-gene-inhibition-property
#7
Kyle K L Phua, Yi Liu, Shun Hui Sim
Nucleic acid induced immunogenicity remains a significant impediment in biomedical therapeutics because the innate immune system is a complex network overlaid with functional redundancies. Herein we report that non-structural protein 1 (NS1), an immune evasion protein derived from influenza A virus, when co-delivered in mRNA format is a potent mRNA transfection enhancer without toxicity. Transfection enhancement is mediated by NS1's effector domain through inhibition of IRF3 and PKR, activators of early anti-viral responses as well as CPSF30, a non immunostimulating protein...
April 12, 2017: Biomaterials
https://www.readbyqxmd.com/read/28417917/minicircle-mediated-gene-delivery-to-canine-and-equine-mesenchymal-stem-cells
#8
Naomie Tidd, Jacob Michelsen, Bryan Hilbert, Jane C Quinn
Gene-directed tissue repair offers the clinician, human or veterinary, the chance to enhance cartilage regeneration and repair at a molecular level. Non-viral plasmid vectors have key biosafety advantages over viral vector systems for regenerative therapies due to their episomal integration however, conventional non-viral vectors can suffer from low transfection efficiency. Our objective was to identify and validate in vitro a novel non-viral gene expression vector that could be utilized for ex vivo and in vivo delivery to stromal-derived mesenchymal stem cells (MSCs)...
April 12, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28412273/efficient-inhibition-of-influenza-a-viral-replication-in-cells-by-deoxyribozymes-delivered-by-nanocomposites
#9
Marina Repkova, Asya Levina, Boris Chelobanov, Zinfer Ismagilov, Natalia Shatskaya, Sergei Baiborodin, Ekaterina Filippova, Natalia Mazurkova, Valentina Zarytova
Nucleic-acid-based drugs are a promising class of novel therapeutics; however, their use in medicine is widely limited because of insufficient delivery into cells. This article proposes a new delivery strategy of nucleic acid fragments into cells as components of TiO2-based nanocomposites. For the first time, unmodified Dz molecules were non-covalently immobilized on TiO2 nanoparticles precovered with polylysine (TiO2•PL) with the formation of (TiO2•PL)•Dz nanocomposites. DNAzymes in the proposed nanocomposites were shown to retain their ability to cleave the RNA target in a cell-free system with the same selectivity as unbound Dz molecules...
April 12, 2017: International Journal of Antimicrobial Agents
https://www.readbyqxmd.com/read/28409504/comparative-study-of-the-transfection-efficiency-of-commonly-used-viral-vectors-in-rhesus-monkey-macaca-mulatta-brains
#10
Shi-Hao Wu, Zhi-Xing Liao, Joshua D Rizak, Na Zheng, Lin-Heng Zhang, Hen Tang, Xiao-Bin He, Yang Wu, Xia-Ping He, Mei-Feng Yang, Zheng-Hui Li, Dong-Dong Qin, Xin-Tian Hu
Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system. In brain research, a series of powerful and novel gene editing technologies are based on these systems. Although many viral vectors are used in rodents, their full application has been limited in non-human primates. To identify viral vectors that can stably and effectively express exogenous genes within non-human primates, eleven commonly used recombinant adeno-associated viral and lentiviral vectors, each carrying a gene to express green or red fluorescence, were injected into the parietal cortex of four rhesus monkeys...
March 18, 2017: Zoological Research
https://www.readbyqxmd.com/read/28397188/brain-targeted-polymers-for-gene-delivery-in-the-treatment-of-brain-diseases
#11
REVIEW
Yifei Lu, Chen Jiang
Gene therapies have become a promising strategy for treating neurological disorders, such as brain cancer and neurodegenerative diseases, with the help of molecular biology interpreting the underlying pathological mechanisms. Successful cellular manipulation against these diseases requires efficient delivery of nucleic acids into brain and further into specific neurons or cancer cells. Compared with viral vectors, non-viral polymeric carriers provide a safer and more flexible way of gene delivery, although suffering from significantly lower transfection efficiency...
April 2017: Topics in Current Chemistry (Journal)
https://www.readbyqxmd.com/read/28396347/serotype-specific-interactions-among-functional-domains-of-dengue-virus-2-non-structural-proteins-ns-5-and-ns3-are-crucial-for-viral-rna-replication
#12
Tadahisa Teramoto, Anuradha Balasubramanian, Kyung H Choi, Radhakrishnan Padmanabhan
Four serotypes of mosquito-borne Dengue virus (DENV), evolved from a common ancestor, are human pathogens of global significance, and there is no vaccine or antiviral drug available. The N-terminal domain of DENV NS5 has guanylyltransferase and methyltransferase (MTase) and the C-terminal region has the polymerase (POL) which are important for 5'-capping and RNA replication. The crystal structure of NS5 showed it as a dimer but the functional evidence for NS5 dimer is lacking. The results of our studies show that the substitution of DENV2 NS5 MTase or POL with that of DENV4 NS5 within DENV2 RNA resulted in severe attenuation of replication in the transfected BHK-21 cells...
April 10, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28393560/evaluating-polyethyleneimine-dna-nanoparticles-mediated-damage-to-cellular-organelles-using-endoplasmic-reticulum-stress-profile
#13
Maryam Dabbaghi, Reza Kazemi Oskuee, Khadijeh Hashemi, Amir Afkhami Goli
Gene therapy has emerged as an influential tool for treating the genetic and specific acquired disorders. Among all kinds of gene delivery systems, the cationic polymer polyethyleneimine (PEI) is considered as a promising non-viral gene delivery vector, although there are still concerns about its magnitude of cytotoxicity. While any cell insult leads to unfolded/misfolded protein accumulation and its consequent unfold protein response, evaluating the expression profile of ER-stress genes would be a sensitive indicator of cell stress...
April 10, 2017: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/28391040/pegylated-composite-nanoparticles-of-plga-and-polyethylenimine-for-safe-and-efficient-delivery-of-pdna-to-lungs
#14
Atul Kolte, Sushilkumar Patil, Pierre Lesimple, John W Hanrahan, Ambikanandan Misra
Achieving stable, efficient and non-toxic pulmonary gene delivery is most challenging requirement for successful gene therapy to lung. Composite nanoparticles (NPs) of the poly(lactic-co-glycolic acid) (PLGA) and cationic polymer polyethyleneimine (PEI) is an efficient alternative to viral and liposomal vectors for the pulmonary delivery of pDNA. NPs with different weight ratios (0-12.5%w/w) of PLGA/PEI were prepared and characterized for size, morphology, surface charge, pDNA loading and in vitro release. The in vitro cell uptake and transfection studies in the CFBE41o-cell line revealed that NPs with 10% w/w PEI were more efficient but they exhibited significant cytotoxicity in MTT assays, challenging the safety of this formulation...
April 6, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/28388487/characterization-of-virus-specific-vesicles-assembled-by-west-nile-virus-non-structural-proteins
#15
Li Yu, Kazuyo Takeda, Yamei Gao
Flavivirus genome encodes seven non-structural proteins (NSPs) and these NSPs are believed to be involved in their genomic RNA replication, of which the mechanism is unclear. We find that West Nile virus (WNV) NSPs were capable of self-assembling membranous vesicles in cells, which are composed of the host endoplasmic reticulum membrane integrated with viral NS1 and NS4A, and possibly NS2A. The vesicles can further organize into replication complex (RC)-associated vesicles which combine both the vesicle and predicted RC components...
June 2017: Virology
https://www.readbyqxmd.com/read/28385616/hydroxyl-versus-permethylated-glycopolymers-as-gene-carriers
#16
Juan Alfonso Redondo, Enrique Martínez-Campos, Rodrigo Navarro, Mónica Pérez-Perrino, Helmut Reinecke, Alberto Gallardo, Guillermo Corrales, Alfonso Fernández-Mayoralas, Carlos Elvira
The main parameters that contribute to non-viral gene delivery are chemical structure and charge distribution. Indeed, saccharide units have been reported to have specific interactions with proteins located in the outer leaflet of the plasma cell membrane that facilitate the cellular internalization of plasmid-DNA vector complexes. In this work, glycopolymers based on statistical copolymers were synthesized through radical copolymerization of a cationic unit, N-ethyl pyrrolidine methacrylamide (EPA), with two styrenic monomers derived from the hydroxylated and permethylated forms of α-glucose...
April 3, 2017: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/28380420/imaging-and-targeted-therapy-of-pancreatic-ductal-adenocarcinoma-using-the-theranostic-sodium-iodide-symporter-nis-gene
#17
Kathrin A Schmohl, Aayush Gupta, Geoffrey K Grünwald, Marija Trajkovic-Arsic, Kathrin Klutz, Rickmer Braren, Markus Schwaiger, Peter J Nelson, Manfred Ogris, Ernst Wagner, Jens T Siveke, Christine Spitzweg
The theranostic sodium iodide symporter (NIS) gene allows detailed molecular imaging of transgene expression and application of therapeutic radionuclides. As a crucial step towards clinical application, we investigated tumor specificity and transfection efficiency of epidermal growth factor receptor (EGFR)-targeted polyplexes as systemic NIS gene delivery vehicles in an advanced genetically engineered mouse model of pancreatic ductal adenocarcinoma (PDAC) that closely reflects human disease. PDAC was induced in mice by pancreas-specific activation of constitutively active KrasG12D and deletion of Trp53...
March 23, 2017: Oncotarget
https://www.readbyqxmd.com/read/28363788/mesenchymal-stem-cell-fate-following-non-viral-gene-transfection-strongly-depends-on-the-choice-of-delivery-vector
#18
T Gonzalez-Fernandez, B N Sathy, C Hobbs, G M Cunniffe, H O McCarthy, N J Dunne, V Nicolosi, F J O'Brien, D J Kelly
Controlling the phenotype of mesenchymal stem cells (MSCs) through the delivery of regulatory genes is a promising strategy in tissue engineering (TE). Essential to effective gene delivery is the choice of gene carrier. Non-viral delivery vectors have been extensively used in TE, however their intrinsic effects on MSC differentiation remain poorly understood. The objective of this study was to investigate the influence of three different classes of non-viral gene delivery vectors: (1) cationic polymers (polyethylenimine, PEI), (2) inorganic nanoparticles (nanohydroxyapatite, nHA) and (3) amphipathic peptides (RALA peptide) on modulating stem cell fate after reporter and therapeutic gene delivery...
March 28, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28362336/biodegradable-poly-amino-ester-with-aromatic-backbone-as-efficient-nonviral-gene-delivery-vectors
#19
Qiang Liu, Rong-Chuan Su, Wen-Jing Yi, Zhi-Gang Zhao
The development of gene delivery vectors with high efficiency and biocompatibility is one of the critical points of gene therapy. Two biodegradable poly(amino ester)s were synthesized via ring-opening polymerization between low molecular weight (LMW) PEI and diepoxide. The molecular weights of poly(amino ester)s were measured by GPC. Agarose gel retardation assays showed that these materials have good DNA-binding ability and can retard the electrophoretic mobility of plasmid DNA (pDNA) at a weight ratio of 1...
March 31, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28352540/promoter-transgene-and-cell-line-effects-in-the-transfection-of-mammalian-cells-using-pdmaema-based-nano-stars
#20
Alexander Raup, Valérie Jérôme, Ruth Freitag, Christopher V Synatschke, Axel H E Müller
Non-viral transfection protocols are typically optimized using standard cells and reporter proteins, potentially underestimating cellular or transgene effects. Here such effects were studied for two human (Jurkat, HEK-293) and two rodent (CHO-K1, L929) cell lines and three fluorescent reporter proteins. Expression of the enhanced green fluorescent protein (EGFP) was studied under the control of the human elongation factor 1 alpha promoter and three viral promoters (SV40, SV40/enhancer, CMV), that of ZsYellow1 (yellow fluorescence) and mCherry (red fluorescence) for the CMV promoter...
September 2016: Biotechnology Reports
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