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https://www.readbyqxmd.com/read/29779176/intranasal-delivery-of-pgdnf-dna-nanoparticles-provides-neuroprotection-in-the-rat-6-hydroxydopamine-model-of-parkinson-s-disease
#1
Amirah E-E Aly, Brendan T Harmon, Linas Padegimas, Ozge Sesenoglu-Laird, Mark J Cooper, Barbara L Waszczak
Glial cell line-derived neurotrophic factor (GDNF) gene therapy could offer a disease-modifying treatment for Parkinson's disease (PD). Here, we report that plasmid DNA nanoparticles (NPs) encoding human GDNF administered intranasally to rats induce transgene expression in the brain and protect dopamine neurons in a model of PD. To first test whether intranasal administration could transfect cells in the brain, rats were sacrificed 1 week after intranasal pGDNF NPs or the naked plasmid. GDNF ELISA revealed significant increases in GDNF expression throughout the brain for both treatments...
May 19, 2018: Molecular Neurobiology
https://www.readbyqxmd.com/read/29778185/serine-105-and-120-are-important-phosphorylation-sites-for-porcine-reproductive-and-respiratory-syndrome-virus-n-protein-function
#2
Yao Chen, Xiulin Xing, Qi Li, Songlin Feng, Xiaoliang Han, Shuyi He, Guihong Zhang
The nucleocapsid (N) protein is the most abundant protein of porcine reproductive and respiratory syndrome virus (PRRSV). It has been shown to be multiphosphorylated. However, the phosphorylation sites are still unknown. In this study, we used liquid chromatography tandem mass spectrometry (LC-MS/MS) to analyze the phosphorylation sites of N protein expressed in Sf9 cells. The results showed that N protein contains two phosphorylation sites. Since N protein can regulate IL-10, which may facilitate PRRSV replication, we constructed four plasmids (pCA-XH-GD, pCA-A105, pCA-A120 and pCA-A105-120) and transfected them into Pig alveolar macrophages (PAMs,3D4/2)...
June 2018: Veterinary Microbiology
https://www.readbyqxmd.com/read/29777445/a-reverse-genetics-system-for-enterovirus-d68-using-human-rna-polymerase-i
#3
Minglei Pan, Shuai Gao, Zhenwei Zhou, Keke Zhang, Sihua Liu, Zhiyun Wang, Tao Wang
Human enterovirus D68 (EV-D68) is a highly contagious virus, which causes respiratory tract infections. However, no effective vaccines are currently available for controlling EV-D68 infection. Here, we developed a reverse genetics system to recover EV-D68 minireplicons and infectious EV-D68 from transfected plasmids using the RNA polymerase I (Pol I) promoter. The EV-D68 minireplicons contained the luciferase reporter gene, which flanked by the non-coding regions of the EV-D68 RNA. The luciferase signals could be detected in cells after transfection and Pol I promoter-mediated luciferase signal was significantly stronger than that mediated by the T7 promoter...
May 17, 2018: Virus Genes
https://www.readbyqxmd.com/read/29775725/anti-hepatitis-c-virus-activity-and-synergistic-effect-of-nymphaea-alba-extracts-and-bioactive-constituents-in-liver-infected-cells
#4
Sidra Rehman, Usman Ali Ashfaq, Bushra Ijaz, Sheikh Riazuddin
BACKGROUND: Without an effective vaccine, hepatitis C virus (HCV) remains a global threat, inflicting 170-300 million carriers worldwide at risk of cirrhosis and hepatocellular carcinoma (HCC). Though various direct acting antivirals have been redeemed the hepatitis C treatment, a few restraints persist including possible side effects, viral resistance emergence, excessive cost which restricts its availability to a common person. HYPOTHESIS: There is no preventive HCV vaccine available today so the discovery of potent antiviral natural flora and their bioactive constituents may help to develop preventive cures against HCV infection...
May 15, 2018: Microbial Pathogenesis
https://www.readbyqxmd.com/read/29773895/crispr-cas9-system-targeting-regulatory-genes-of-hiv-1-inhibits-viral-replication-in-infected-t-cell-cultures
#5
Youdiil Ophinni, Mari Inoue, Tomohiro Kotaki, Masanori Kameoka
The CRISPR/Cas9 system provides a novel and promising tool for editing the HIV-1 proviral genome. We designed RNA-guided CRISPR/Cas9 targeting the HIV-1 regulatory genes tat and rev with guide RNAs (gRNA) selected from each gene based on CRISPR specificity and sequence conservation across six major HIV-1 subtypes. Each gRNA was cloned into lentiCRISPRv2 before co-transfection to create a lentiviral vector and transduction into target cells. CRISPR/Cas9 transduction into 293 T and HeLa cells stably expressing Tat and Rev proteins successfully abolished the expression of each protein relative to that in non-transduced and gRNA-absent vector-transduced cells...
May 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29766760/osteoinduction-by-ex-vivo-bone-morphogenetic-protein-gene-delivery-is-independent-of-cell-type
#6
Loek Loozen, Moyo C Kruyt, Angela Vandersteen, Angela H M Kragten, Michiel Croes, F Cumhur Oner, Jacqueline Alblas
Ex vivo non-viral gene delivery of bone-inductive factors has the potential to heal bone defects. Due to their inherent role in new bone formation, multipotent stromal cells (MSCs) have been studied as the primary target cell for gene delivery in a preclinical setting. The relative contribution of autocrine and paracrine mechanisms, and the need of osteogenic cells, remains unclear. This study investigates the contribution of MSCs as producer of transgenic BMPs and to what extent the seeded MSCs participate in actual osteogenesis...
May 16, 2018: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/29755438/caspase-3-inhibition-attenuates-the-cytopathic-effects-of-ev71-infection
#7
Fengmei Song, Xiaoyan Yu, Ting Zhong, Zengyan Wang, Xiangling Meng, Zhaolong Li, Shuxia Zhang, Wenbo Huo, Xin Liu, Yahong Zhang, Wenyan Zhang, Jinghua Yu
Previous studies demonstrate that human enterovirus 71 (EV71), a primary causative agent for hand, foot, and mouth disease, activates caspase-3 through the non-structural viral 3C protein to induce host cell apoptosis; however, until now it was unclear how 3C activates caspase-3 and how caspase-3 activation affects viral production. Our results demonstrate that 3C binds caspase-8 and caspase-9 but does not directly bind caspase-3 to activate them, and that the proteolytic activity of 3C is required by the activation of caspase-8, caspase-9, and caspase-3...
2018: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29745740/co-delivery-of-gambogenic-acid-and-vegf-sirna-with-anionic-liposome-and-polyethylenimine-complexes-to-hepg2-cells
#8
Qiongfang Yu, Bian Zhang, Yali Zhou, Qin Ge, Chang Jiali, Yunna Chen, Kaiqi Zhang, Daiyin Peng, Weidong Chen
Background and Objective The combination of two or more different mechanisms of drugs in the treatment of cancer has become one of the effective methods. The purpose of this study was to successfully prepare a non-viral delivery system that could efficiently co-delivery siRNA and gambogenic acid (GNA) to improve the anti-cancer efficiency in HepG2 cells. Methods The delivery system was prepared by a two-step method. Firstly, the GNA anionic liposome took shape by a solvent evaporation method, and then the liposome was bound to the PEI/siRNA complex by electrostatic interaction to form the final carrier system (lipopolyplexes)...
May 10, 2018: Journal of Liposome Research
https://www.readbyqxmd.com/read/29728593/photoluminescent-cationic-carbon-dots-as-efficient-non-viral-delivery-of-plasmid-sox9-and-chondrogenesis-of-fibroblasts
#9
Xia Cao, Jianping Wang, Wenwen Deng, Jingjing Chen, Yan Wang, Jie Zhou, Pan Du, Wenqian Xu, Qiang Wang, Qilong Wang, Qingtong Yu, Myron Spector, Jiangnan Yu, Ximing Xu
With the increasing demand for higher gene carrier performance, a multifunctional vector could immensely simplify gene delivery for disease treatment; nevertheless, the current non- viral vectors lack self-tracking ability. Here, a type of novel, dual-functional cationic carbon dots (CDs), produced through one-step, microwave-assisted pyrolysis of arginine and glucose, have been utilized as both a self-imaging agent and a non-viral gene vector for chondrogenesis from fibroblasts. The cationic CDs could condense the model gene plasmid SOX9 (pSOX9) to form ultra-small (10-30 nm) nanoparticles which possessed several favorable properties, including high solubility, tunable fluorescence, high yield, low cytotoxicity and outstanding biocompatibility...
May 4, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29721934/optimizing-non-viral-gene-therapy-vectors-for-delivery-to-photoreceptors-and-retinal-pigment-epithelial-cells
#10
Rahel Zulliger, Jamie N Watson, Muayyad R Al-Ubaidi, Linas Padegimas, Ozge Sesenoglu-Laird, Mark J Cooper, Muna I Naash
Considerable progress has been made in the design and delivery of non-viral gene therapy vectors, but, like their viral counterparts, therapeutic levels of transgenes have not met the requirements for successful clinical applications so far. The biggest advantage of polymer-based nanoparticle vectors is the ease with which they can be modified to increase their ability to penetrate the cell membrane and target specific cells by simply changing the formulation of the nanoparticle compaction. We took advantage of this characteristic to improve transfection rates of our particles to meet the transgene levels which will be needed for future treatment of patients...
2018: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/29712427/beyond-gene-transfection-with-methacrylate-based-polyplexes-the-influence-of-the-amino-substitution-pattern
#11
Ulrich S Schubert, Tanja Bus, Martin Reifarth, Johannes C Brendel, Stephanie Hoeppener, Anja Traeger, Anne-Kristin Trützschler
Methacrylate-based polymers represent promising non-viral gene delivery vectors, since they offer a large variety of polymer architectures and functionalities, which are beneficial for specific demands in gene delivery. In combination with controlled radical polymerization techniques, such as the reversible addition-fragmentation chain transfer polymerization, the synthesis of well-defined polymers is possible. In this study we prepared a library of defined linear polymers based on (2-aminoethyl)-methacrylate (AEMA), N-methyl-(2-aminoethyl)-methacrylate (MAEMA) and N,N-dimethyl-(2-aminoethyl)-methacrylate (DMAEMA) monomers, bearing pendant primary, secondary and tertiary amino groups, and investigated the influence of the substitution pattern on their gene delivery ability...
May 1, 2018: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/29693099/deciphering-the-role-of-substrate-stiffness-in-enhancing-the-internalization-efficiency-of-plasmid-dna-in-stem-cells-using-lipid-based-nanocarriers
#12
Saman Modaresi, Settimio Pacelli, Jonathan Whitlow, Arghya Paul
This study investigates the role of substrate stiffness in the non-viral transfection of human adipose-derived stem cells (hASCs) with the aim to maximize the hASC expression of vascular endothelial growth factor (VEGF). The results confirm the direct effect of substrate stiffness in regulating cytoskeletal remodeling and corresponding plasmid internalization.
April 25, 2018: Nanoscale
https://www.readbyqxmd.com/read/29689167/tailoring-the-supramolecular-structure-of-guanidinylated-pullulan-toward-enhanced-genetic-photodynamic-therapy
#13
Jie Zhou, Aisha Roshan Mohamed Wali, Shengnan Ma, Yiyan He, Dong Yue, James Zhenggui Tang, Zhongwei Gu
In the progress of designing a gene carrier system, what is urgently needed is a balance of excellent safety and satisfactory efficiency. Herein, a straightforward and versatile synthesis of a cationic guanidine-decorated dendronized pullulan (OGG3P) for efficient genetic photodynamic therapy was proposed. OGG3P was able to block the mobility of DNA from a weight ratio of 2. However, G3P lacking guanidine residues could not block DNA migration until at a weight ratio of 15, revealing guanidination could facilitate DNA condensation via specific guanidinium-phosphate interactions...
April 24, 2018: Biomacromolecules
https://www.readbyqxmd.com/read/29687032/enhanced-expression-of-anti-cd19-chimeric-antigen-receptor-in-piggybac-transposon-engineered-t-cells
#14
Daisuke Morita, Nobuhiro Nishio, Shoji Saito, Miyuki Tanaka, Nozomu Kawashima, Yusuke Okuno, Satoshi Suzuki, Kazuyuki Matsuda, Yasuhiro Maeda, Matthew H Wilson, Gianpietro Dotti, Cliona M Rooney, Yoshiyuki Takahashi, Yozo Nakazawa
Adoptive T cell therapy using chimeric antigen receptor (CAR)-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a piggyBac transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon vector and combining irradiated activated T cells (ATCs) as feeder cells and virus-specific T cell receptor (TCR) stimulation, we achieved 51...
March 16, 2018: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/29658818/a-long-non-coding-sineup-rna-boosts-semi-stable-production-of-fully-human-monoclonal-antibodies-in-hek293e-cells
#15
Emanuele Sasso, Debora Latino, Guendalina Froechlich, Mariangela Succoio, Margherita Passariello, Claudia De Lorenzo, Alfredo Nicosia, Nicola Zambrano
Use of monoclonal antibodies is emerging as a highly promising and fast-developing scenario for innovative treatment of viral, autoimmune and tumour diseases. The search for diagnostic and therapeutic antibodies currently depends on in vitro screening approaches, such as phage and yeast display technologies. Antibody production still represents a critical step for preclinical and clinical evaluations. Accordingly, improving production of monoclonal antibodies represents an opportunity, to facilitate downstream target validations...
April 16, 2018: MAbs
https://www.readbyqxmd.com/read/29618647/identification-of-the-rna-pseudoknot-within-the-3-end-of-prrsv-genome-as-pathogen-associated-molecular-pattern-to-activate-antiviral-signaling-via-rig-i-and-tlr3
#16
Sha Xie, Xin-Xin Chen, Songlin Qiao, Rui Li, Yangang Sun, Shuangfei Xia, Lin-Jian Wang, Xuegang Luo, Ruiguang Deng, En-Min Zhou, Gai-Ping Zhang
Once infected by viruses, cells can detect pathogen associated molecular patterns (PAMPs) on viral nucleic acid by host pattern recognition receptors (PRRs) to initiate the antiviral response. Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome (PRRS), characterized by reproductive failure in sows and respiratory diseases in pigs of different ages. To date, the sensing mechanism of PRRSV has not been elucidated. Here, we reported that the pseudoknot region residing in the 3' untranslated regions (UTR) of PRRSV genome, which has been proposed to regulate RNA synthesis and virus replication, was sensed as non-self by retinoic acid-inducible gene I (RIG-I) and Toll like receptor (TLR3) and strongly induced type I IFNs and interferon-stimulated genes (ISGs) in porcine alveolar macrophages (PAMs)...
April 4, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29615605/metal-organic-frameworks-for-precise-inclusion-of-single-stranded-dna-and-transfection-in-immune-cells
#17
Shuang Peng, Binglin Bie, Yangzesheng Sun, Min Liu, Hengjiang Cong, Wentao Zhou, Yucong Xia, Heng Tang, Hexiang Deng, Xiang Zhou
Effective transfection of genetic molecules such as DNA usually relies on vectors that can reversibly uptake and release these molecules, and protect them from digestion by nuclease. Non-viral vectors meeting these requirements are rare due to the lack of specific interactions with DNA. Here, we design a series of four isoreticular metal-organic frameworks (Ni-IRMOF-74-II to -V) with progressively tuned pore size from 2.2 to 4.2 nm to precisely include single-stranded DNA (ssDNA, 11-53 nt), and to achieve reversible interaction between MOFs and ssDNA...
April 3, 2018: Nature Communications
https://www.readbyqxmd.com/read/29611333/real-time-imaging-of-dynamic-cell-reprogramming-with-nanosensors
#18
Christian Wiraja, David C Yeo, Khek-Chian Tham, Sharon W T Chew, Xinhong Lim, Chenjie Xu
Cellular reprogramming, the process by which somatic cells regain pluripotency, is relevant in many disease modeling, therapeutic, and drug discovery applications. Molecular evaluation of reprogramming (e.g., polymerase chain reaction, immunostaining) is typically disruptive, and only provides snapshots of phenotypic traits. Gene reporter constructs facilitate live-cell evaluation but is labor intensive and may risk insertional mutagenesis during viral transfection. Herein, the utilization of a non-integrative nanosensor is demonstrated to visualize key reprogramming events in situ within live cells...
April 3, 2018: Small
https://www.readbyqxmd.com/read/29593359/magnetic-nanoparticle-mediated-gene-therapy-to-induce-fas-apoptosis-pathway-in-breast-cancer
#19
Harun Basoglu, Beyza Goncu, Fahri Akbas
CD95 (Fas) is a complex integral protein that can be expressed in many cells. It induces apoptosis when interacted with its ligand CD95L (FasL). However, cancer cells are resistant to CD95-induced apoptosis because of the changes in death domain (DD) of CD95 (procaspase-8 and c-Flip). In this study, magnetic nanoparticles and lipid-based gene transfection methods were performed to provide active Fas expression in breast cancer cells. Plasmid DNA (pDNA), which can express both human Fas and GFP, was transfected to MCF-7 breast cancer cells...
March 29, 2018: Cancer Gene Therapy
https://www.readbyqxmd.com/read/29590755/bioinspired-star-shaped-poly-l-lysine-polypeptides-efficient-polymeric-nanocarriers-for-the-delivery-of-dna-to-mesenchymal-stem-cells
#20
David P Walsh, Robert D Murphy, Angela Panarella, Rosanne Marie Raftery, Brenton Cavanagh, Jeremy Charles Simpson, Fergal J O'Brien, Andreas Heise, Sally-Ann Cryan
The field of tissue engineering is increasingly recognizing that gene therapy can be employed for modulating in vivo cellular response thereby guiding tissue regeneration. However, the field lacks a versatile and biocompatible gene delivery platform capable of efficiently delivering transgenes to mesenchymal stem cells (MSCs), a cell type refractory to transfection. Herein, we describe the extensive and systematic exploration of three architectural variations of star-shaped poly(L-lysine) polypeptide (star-PLL) with varying number and length of poly(L-Lysine) arms as potential non-viral gene delivery vectors for MSCs...
March 28, 2018: Molecular Pharmaceutics
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