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https://www.readbyqxmd.com/read/28632911/polymer-functionalized-gold-nanoparticles-as-non-viral-gene-delivery-reagents
#1
M Mar Encabo-Berzosa, Maria Sancho-Albero, Victor Sebastian, Silvia Irusta, Manuel Arruebo, Jesus Santamaria, Pilar Martín Duque
BACKGROUND: In this study we investigated the ability of PEG functionalized gold nanoparticles as non-viral vectors in the transfection of different cell lines, comparing them with commercial lipoplexes. METHODS: Positively charged gold nanoparticles were synthesized using PEI as reducing and stabilizer agent and its cytotoxicity reduced by its functionalization with PEG. We bound the nanoparticles to three plasmids with different sizes (4-40 kpb). The vector internalization was evaluated by confocal and electronic microscopy...
June 20, 2017: Journal of Gene Medicine
https://www.readbyqxmd.com/read/28629980/polymer-nanoassemblies-with-hydrophobic-pendant-groups-in-the-core-induce-false-positive-sirna-transfection-in-luciferase-reporter-assays
#2
Steven Rheiner, Derek Reichel, Piotr Rychahou, Tadahide Izumi, Hsin-Sheng Yang, Younsoo Bae
Poly(ethylene glycol)-conjugated polyethylenimine (PEG-PEI) is a widely studied cationic polymer used to develop non-viral vectors for siRNA therapy of genetic disorders including cancer. Cell lines stably expressing luciferase reporter protein typically evaluate the transfection efficacy of siRNA/PEG-PEI complexes, however recent findings revealed that PEG-PEI can reduce luciferase expression independent of siRNA. This study elucidates a cause of the false positive effect in luciferase assays by using polymer nanoassemblies (PNAs) made from PEG, PEI, poly-(L-lysine) (PLL), palmitate (PAL), and deoxycholate (DOC): PEG-PEI (2P), PEG-PEI-PAL (3P), PEG-PLL (2P'), PEG-PLL-PAL (3P'), and PEG-PEI-DOC (2PD)...
June 16, 2017: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/28629080/efficient-megalin-targeted-delivery-to-renal-proximal-tubular-cells-mediated-by-modified-polymyxin-b-polyethylenimine-based-nano-gene-carriers
#3
Fatemeh Oroojalian, Ali Hossein Rezayan, Faramarz Mehrnejad, Azadeh Hashem Nia, Wayne Thomas Shier, Khalil Abnous, Mohammad Ramezani
Non-viral vectors have attracted great interest, as they are simple to prepare, easy to modify and relatively safe, compared to viral vectors. Kidney-targeted gene delivery systems depict a promising technology to improve drug efficacy in renal diseases treatments. In order to develop a novel kidney-targeted gene delivery system, we synthesized polyamine-PEI conjugates using polymyxin B as ligand and investigated their potential targeting efficiency. After grafting either PEI25 kDa or PEI10 kDa with polymyxin B through amidation reaction, the modified-polymyxin-PEI/DNA-nanoplexes were produced via electrostatic attraction between the cationic polymers and EGFP plasmid...
October 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28624210/low-dose-gene-therapy-for-murine-pku-using-episomal-naked-dna-vectors-expressing-pah-from-its-endogenous-liver-promoter
#4
Hiu Man Grisch-Chan, Andrea Schlegel, Tanja Scherer, Gabriella Allegri, Raphael Heidelberger, Panagiota Tsikrika, Marco Schmeer, Martin Schleef, Cary O Harding, Johannes Häberle, Beat Thöny
Limited duration of transgene expression, insertional mutagenesis, and size limitations for transgene cassettes pose challenges and risk factors for many gene therapy vectors. Here, we report on physiological expression of liver phenylalanine hydroxylase (PAH) by delivery of naked DNA/minicircle (MC)-based vectors for correction of homozygous enu2 mice, a model of human phenylketonuria (PKU). Because MC vectors lack a defined size limit, we constructed a MC vector expressing a codon-optimized murine Pah cDNA that includes a truncated intron and is under the transcriptional control of a 3...
June 16, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28618452/murine-melanoma-cells-incomplete-reprogramming-using-non-viral-vector
#5
D A D Câmara, A S Porcacchia, A S Costa, R A Azevedo, I Kerkis
OBJECTIVES: The reprogramming of cancer cells into induced pluripotent stem cells or less aggressive cancer cells can provide a modern platform to study cancer-related genes and their interactions with cell environment before and after reprogramming. Herein, we aimed to investigate the reprogramming capacity of murine melanoma B16F10 cells. MATERIALS AND METHODS: The B16F10 was transfected using non-viral circular DNA plasmid containing the genes Sox-2, Oct4, Nanog, Lin28 and green fluorescent protein (GFP)...
June 15, 2017: Cell Proliferation
https://www.readbyqxmd.com/read/28618303/transcriptomic-response-of-the-insect-vector-peregrinus-maidis-to-maize-mosaic-rhabdovirus-and-identification-of-conserved-responses-to-propagative-viruses-in-hopper-vectors
#6
Kathleen M Martin, Karen Barandoc-Alviar, Derek J Schneweis, Catherine L Stewart, Dorith Rotenberg, Anna E Whitfield
Maize mosaic virus (MMV) is a plant-pathogenic rhabdovirus that is transmitted by the corn planthopper, Peregrinus maidis, in a propagative manner. P. maidis supports long-term MMV infections with no negative effects on insect performance. To elucidate whole-body transcriptome responses to virus infection, RNA-Seq was used to examine differential gene expression of virus-infected adult insects, and libraries were prepared from replicated groups of virus-exposed insects and non-exposed insects. From the 68,003 de novo-assembled transcripts, 144 were differentially-expressed (DE) during viral infection with comparable numbers up- and down-regulated...
June 12, 2017: Virology
https://www.readbyqxmd.com/read/28609784/genome-wide-profiling-of-s-mar-based-replicon-contact-sites
#7
Claudia Hagedorn, Andreas Gogol-Döring, Sabrina Schreiber, Jörg T Epplen, Hans J Lipps
Autonomously replicating vectors represent a simple and versatile model system for genetic modifications, but their localization in the nucleus and effect on endogenous gene expression is largely unknown. Using circular chromosome conformation capture we mapped genomic contact sites of S/MAR-based replicons in HeLa cells. The influence of cis-active sequences on genomic localization was assessed using replicons containing either an insulator sequence or an intron. While the original and the insulator-containing replicons displayed distinct contact sites, the intron-containing replicon showed a rather broad genomic contact pattern...
June 13, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28605938/a-guide-to-approaching-regulatory-considerations-for-lentiviral-mediated-gene-therapies
#8
Michael White, Roger Whittaker, Elizabeth Ann Stoll
Lentiviral vectors are increasingly the gene transfer tool of choice for gene or cell therapies, with multiple clinical investigations showing promise for this viral vector in terms of both safety and efficacy. The third-generation vector system is well-characterized, effectively delivers genetic material and maintains long-term stable expression in target cells, delivers larger amounts of genetic material than other methods, is non-pathogenic and does not cause an inflammatory response in the recipient. This report aims to help academic scientists and regulatory managers negotiate the governance framework to achieve successful translation of a lentiviral vector-based gene therapy...
June 12, 2017: Human Gene Therapy Methods
https://www.readbyqxmd.com/read/28603974/evolution-of-new-bolaliposomes-using-novel-%C3%AF-tocopheryl-succinate-based-cationic-lipid-and-1-12-disubstituted-dodecane-based-bolaamphiphile-for-efficient-gene-delivery
#9
Mallikarjun Gosangi, Hithavani Rapaka, Venkatesh Ravula, Srilakshmi Venkata Patri
Non-viral lipid based vectors are promising transporting systems for the intracellular delivery of therapeutic gene sequences and directly influence the success of gene delivery. However, the associated drawbacks like lower transfection, toxicity and targetability with which demands further improvisation. Towards this goal, herein, we report a novel lipid formulation by mixing two distinct cationic surfactants such as tocopheryl succinate based cationic lipid and 1, 12 dodecane based bolaamphiphile and is proved as good transfection reagent with its competing potential with "golden" standard, Lipofectamine 3000 (L3K)...
June 12, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28600130/highly-antibacterial-polypeptide-based-amphiphilic-copolymers-as-multifunctional-non-viral-vectors-for-enhanced-intracellular-sirna-delivery-and-anti-infection
#10
Li Zhou, Yuewei Xi, Meng Yu, Min Wang, Yi Guo, Peng Li, Peter X Ma, Bo Lei
For next-generation regenerative medicine and cancer therapy applications, the development of multifunctional biodegradable biomaterials with good antibacterial activity, biocompatibility and gene delivery capability is showing increased interests. As a natural cationic polypeptide, poly(ε-L-lysine) (EPL) has been as attractive antibacterial agent and non-viral gene vectors, however, its gene therapy is still limited probably due to the cytotoxicity, low stability in physiological environment and poor transfection efficiency...
June 6, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28597662/chitosan-hyaluronic-acid-nanoparticles-rational-design-revisited-for-rna-delivery
#11
Enrique Lallana, Julio M Rios de la Rosa, Annalisa Tirella, Maria Pelliccia, Arianna Gennari, Ian J Stratford, Sanyogitta Puri, Marianne Ashford, Nicola Tirelli
Chitosan/hyaluronic acid (HA) nanoparticles can be used to deliver an RNA/DNA cargo to cells overexpressing HA receptors such as CD44. For these systems, unequivocal links have not been established yet between chitosan macromolecular (molecular weight; degree of deacetylation, i.e., charge density) and nanoparticle variables (complexation strength, i.e., stability; nucleic acid protection; internalization rate) on one hand, and transfection efficiency on the other hand. Here, we have focused on the role of avidity on transfection efficiency in the CD44-expressing HCT-116 as a cellular model; we have employed two differently sized payloads (a large luciferase-encoding mRNA and a much smaller anti-Luc siRNA), and a small library of chitosans (variable molecular weight and degree of deactylation)...
June 19, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28596158/molecular-imaging-of-human-embryonic-stem-cells-stably-expressing-human-pet-reporter-genes-after-zinc-finger-nucleases-mediated-genome-editing
#12
Esther Wolfs, Bryan Holvoet, Laura Ordovas, Natacha Breuls, Nicky Helsen, Matthias Schönberger, Susanna Raitano, Tom Struys, Bert Vanbilloen, Cindy Casteels, Maurilio Sampaolesi, Koen Van Laere, Ivo Lambrichts, Catherine M Verfaillie, Christophe M Deroose
Molecular imaging is indispensable for determining the fate and persistence of engrafted stem cells. Standard strategies for transgene induction involve the use of viral vectors prone to silencing and insertional mutagenesis or the use of non-human genes. We used zinc finger nucleases (ZFN) to induce stable expression of human imaging reporter genes into the safe harbor locus adeno-associated virus integration site 1 (AAVS1) in human embryonic stem cells (ESC). Plasmids were generated carrying reporter genes for fluorescence, bioluminescence imaging (BLI), and human positron emission tomography (PET) reporter genes...
June 8, 2017: Journal of Nuclear Medicine: Official Publication, Society of Nuclear Medicine
https://www.readbyqxmd.com/read/28595562/non-viral-delivery-systems-for-breast-cancer-gene-therapy
#13
Golnaz Vaseghi, Laleh Rafiee, Shaghayegh Haghjooy Javanmard
INTRODUCTION: The ever-evolving field of gene therapy promises several innovative treatments for cancer. Advances in genetic modification of tumor cells and micro environment have led to the development of more effective therapeutic strategies with fewer side effects. MATERIAL & METHODS: The development of effective delivery system challenges, remains. Non-viral vectors are interesting due to their bio-safety and their ability to transfer different types of nucleic acids...
June 6, 2017: Current Gene Therapy
https://www.readbyqxmd.com/read/28593513/gene-therapy-with-helper-dependent-adenoviral-vectors-lessons-from-studies-in-large-animal-models
#14
REVIEW
Nicola Brunetti-Pierri, Philip Ng
Helper-dependent adenoviral vectors (HDAd) are deleted of all viral genes and they can efficiently transduce a wide variety of dividing and non-dividing cells to mediate high transgene expression levels. Unlike early generation adenoviral vectors, the absence of viral genes in HDAd results in long-term transgene expression without chronic toxicity and permits a large cloning capacity of 36 kb. Moreover, HDAd genomes exist extra-chromosomally thus minimizing the risks of germline transmission and insertional mutagenesis...
June 7, 2017: Virus Genes
https://www.readbyqxmd.com/read/28592531/use-of-a-recombinant-gamma-2-herpesvirus-vaccine-vector-against-dengue-virus-in-rhesus-monkeys
#15
Georg F Bischof, Diogo M Magnani, Michael Ricciardi, Young C Shin, Aline Domingues, Varian K Bailey, Lucas Gonzalez-Nieto, Eva G Rakasz, David I Watkins, Ronald C Desrosiers
Research on vaccine approaches that can provide long-term protection against dengue virus infection is needed. Here we describe the construction, immunogenicity, and preliminary information on protective capacity of recombinant, replication-competent rhesus monkey rhadinovirus (RRV), a persisting herpesvirus. One RRV construct expressed the non-structural protein 5 (NS5) while a second recombinant expressed a soluble variant of the E protein (E85) of dengue virus 2 (DENV2). Four rhesus macaques received a single vaccination with a mixture of both recombinant RRVs and were subsequently challenged 19 weeks later with 1 x 10(5) PFU of DENV2...
June 7, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28592330/state-of-play-and-clinical-prospects-of-antibody-gene-transfer
#16
REVIEW
Kevin Hollevoet, Paul J Declerck
Recombinant monoclonal antibodies (mAbs) are one of today's most successful therapeutic classes in inflammatory diseases and oncology. A wider accessibility and implementation, however, is hampered by the high product cost and prolonged need for frequent administration. The surge in more effective mAb combination therapies further adds to the costs and risk of toxicity. To address these issues, antibody gene transfer seeks to administer to patients the mAb-encoding nucleotide sequence, rather than the mAb protein...
June 7, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/28589078/recent-clinical-trials-of-cancer-immunogene-therapy-in-companion-animals
#17
REVIEW
Liliana M E Finocchiaro, Gerardo C Glikin
This mini-review presents the results of veterinary clinical trials on immunogene therapy published from 2014 to 2016. A variety of tumors, among them melanoma (canine and equine), mastocytoma (canine), mammary adenocarcinoma (canine) and fibrosarcoma (feline) were treated by using diverse strategies. Non-viral vectors were usually employed to transfer genes of cytokines, suicide enzymes and/or tumor associated antigens. In general terms, minor or no adverse collateral effects were related to these procedures, and treated patients frequently improved their conditions (better quality of life, delayed or suppressed recurrence or metastatic spread, increased survival)...
May 20, 2017: World Journal of Experimental Medicine
https://www.readbyqxmd.com/read/28588449/long-term-assessment-of-aav-mediated-zinc-finger-nuclease-expression-in-the-mouse-brain
#18
Muzna Zahur, Johan Tolö, Mathias Bähr, Sebastian Kügler
Gene editing tools like TALENs, ZFNs and Crispr/Cas now offer unprecedented opportunities for targeted genetic manipulations in virtually all species. Most of the recent research in this area has concentrated on manipulation of the genome in isolated cells, which then give rise to transgenic animals or modified stem cell lines. Much less is known about applicability of genetic scissors in terminally differentiated, non-dividing cells like neurons of the adult brain. We addressed this question by expression of a pair of ZFNs targeting the murine cathepsin D gene in CNS neurons by means of an optimized AAV viral vector...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28587865/impact-of-vat-resistance-in-melon-on-viral-epidemics-and-genetic-structure-of-virus-populations
#19
Schoeny Alexandra, Desbiez Cécile, Millot Pauline, Wipf-Scheibel Catherine, Nozeran Karine, Gognalons Patrick, Lecoq Hervé, Boissot Nathalie
Cultivar choice is at the heart of cropping systems and resistant cultivars should be at the heart of disease management strategies whenever available. They are the easiest, most efficient and environmentally friendly way of combating viral diseases at the farm level. Among the melon genetic resources, Vat is a unique gene conferring resistance to both the melon aphid Aphis gossypii and the viruses it carries. The 'virus side' of this pleiotropic phenotype is seldom regarded as an asset for virus control. Indeed, the effect of Vat on virus epidemics in the field is expected to vary according to the composition of aphid populations in the environment and long-term studies are needed to draw a correct trend...
June 3, 2017: Virus Research
https://www.readbyqxmd.com/read/28587300/the-5-and-3-untranslated-regions-of-the-flaviviral-genome
#20
REVIEW
Wy Ching Ng, Ruben Soto-Acosta, Shelton S Bradrick, Mariano A Garcia-Blanco, Eng Eong Ooi
Flaviviruses are enveloped arthropod-borne viruses with a single-stranded, positive-sense RNA genome that can cause serious illness in humans and animals. The 11 kb 5' capped RNA genome consists of a single open reading frame (ORF), and is flanked by 5' and 3' untranslated regions (UTR). The ORF is a polyprotein that is processed into three structural and seven non-structural proteins. The UTRs have been shown to be important for viral replication and immune modulation. Both of these regions consist of elements that are essential for genome cyclization, resulting in initiation of RNA synthesis...
June 6, 2017: Viruses
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