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https://www.readbyqxmd.com/read/28528119/autophagy-plays-a-dual-role-during-intracellular-sirna-delivery-by-lipoplex-and-polyplex-nanoparticles
#1
Wen Song, Zhiwei Ma, Yumei Zhang, Chuanxu Yang
Growing evidence indicates that autophagy plays a vital role during the intracellular DNA delivery mediated by lipoplex and polyplex nanoparticles. However, autophagy in intracellular siRNA delivery has not been well understood. In this study, lipofectamine 2000 and chitosan were used to formulate lipoplex and polyplex with siRNA for systematically investigating the interplay between siRNA delivery and autophagy. After transfection of H1299 cells with lipoplex and polyplex, the number of autophagic vacuoles was increased significantly indicated by the accumulation of monodansylcadaverine (MDC) staining...
May 17, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28515301/the-influenza-a-virus-ns1-protein-promotes-efficient-nuclear-export-of-unspliced-viral-m1-mrna
#2
Carina F Pereira, Eliot K C Read, Helen M Wise, Maria J Amorim, Paul Digard
Influenza A virus mRNAs are transcribed by the viral RNA-dependent RNA polymerase in the cell nucleus before being exported to the cytoplasm for translation. Segment 7 produces two major transcripts: an unspliced mRNA that encodes the M1 matrix protein and a spliced transcript that encodes the M2 ion channel. Export of both mRNAs is dependent on the cellular NXF1/TAP pathway but it is unclear how they are recruited to the export machinery or how the intron-containing but unspliced M1 mRNA bypasses the normal quality control checkpoints...
May 17, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28509532/nanodiamonds-mediate-oral-delivery-of-proteins-for-stem-cell-activation-and-intestinal-remodeling-in-drosophila
#3
Xingjie Hu, Xiaojiao Li, Min Yin, Ping Li, Ping Huang, Lihua Wang, Yiguo Jiang, Hui Wang, Nan Chen, Chunhai Fan, Haiyun Song
Introduction of exogenous bio-macromolecules into living systems is of great interest in genome editing, cancer immunotherapy and stem cell reprogramming. Whereas current strategies generally depend on nucleic acids transfection, direct delivery of functional proteins that provides enhanced specificity, increased safety, fast and temporal regulation is highly desirable. Nevertheless, intracellular delivery of intact and bioactive proteins, especially in vivo, remains poorly explored. In this study, we developed a nanodiamonds (NDs)-based protein delivery system in cultured cells and in Drosophila, which showed high adsorption, high efficiency and effective cytosolic release of fully functional proteins...
May 16, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28504656/effect-of-il-1%C3%AE-tnf-%C3%AE-and-igf-1-on-trans-endothelial-passage-of-synthetic-vectors-through-an-in-vitro-vascular-endothelial-barrier-of-striated-muscle
#4
J-P Gomez, C Gonçalves, C Pichon, P Midoux
When administrated in the blood circulation, plasmid DNA (pDNA) complexed with synthetic vectors must pass through a vascular endothelium to transfect underlying tissues. Under inflammatory condition, cytokines effects can modify the endothelium integrity. Here, the trans-endothelial passage (TEP) of DNA complexes including polyplexes, lipoplexes and lipopolyplexes was investigated in the presence of TNF-α-, IL-1β or IGF-1. The experiments were performed by using an in vitro model comprising a monolayer of mouse cardiac endothelial cells (MCEC) seeded on a trans-well insert and the transfection of C2C12 myoblasts cultured on the lower chamber as read out of TEP...
May 15, 2017: Gene Therapy
https://www.readbyqxmd.com/read/28501631/plasmid-transfection-in-bovine-cells-optimization-using-a-realtime-monitoring-of-green-fluorescent-protein-and-effect-on-gene-reporter-assay
#5
J S Osorio, M Bionaz
Gene reporter technology (GRT) has opened several new avenues for monitoring biological events including the activation of transcription factors, which are central to the study of nutrigenomics. However, this technology relies heavily on the insertion of foreign plasmid DNA into the nuclei of cells (i.e., transfection), which can be very challenging and highly variable among cell types. The objective of this study was to investigate the optimal conditions to generate reliable GRT assay data on bovine immortalized cell lines, Madin Darby Bovine Kidney (MDBK) and bovine mammary epithelial alveolar (MACT) cells...
May 10, 2017: Gene
https://www.readbyqxmd.com/read/28496379/self-organization-of-nucleic-acids-in-lipid-constructs
#6
Minjee Kang, Hojun Kim, Cecilia Leal
Lipids and nucleic acids (NAs) can hierarchically self-organize into a variety of nanostructures of increasingly complex geometries such as the 1D lamellar, 2D hexagonal, and 3D bicontinuous cubic phases. The diversity and complexity of those lipid-NA assemblies are interesting from a fundamental perspective as well as being relevant to the performance in gene delivery and gene silencing applications. The finding that not only the chemical make of the lipid-NA constructs, but their actual supramolecular organization, affects their gene transfection and silencing efficiencies has inspired physicists, chemists, and engineers to this field of research...
December 2016: Current Opinion in Colloid & Interface Science
https://www.readbyqxmd.com/read/28476488/improving-pertuzumab-production-by-gene-optimization-and-proper-signal-peptide-selection
#7
Amin Ramezani, Elham Mahmoudi Maymand, Mahsa Yazdanpanah-Samani, Ahmad Hosseini, Fatemeh Sadat Toghraie, Abbas Ghaderi
Using proper signal peptide and codon optimization are important factors that must be considered when designing the vector to increase protein expression in Chinese Hamster Ovary (CHO) cells. The aim of the present study is to investigate how to enhance Pertuzumab production through heavy and light chain coding gene optimization and proper signal peptide selection. First, CHO-K1 cells were transiently transfected with whole-antibody-gene-optimized, variable-regions-optimized and non-optimized constructs and then we employed five different signal peptides to improve the secretion efficiency of Pertuzumab...
May 3, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28463994/influence-of-nanoparticle-mediated-transfection-on-proliferation-of-primary-immune-cells-in-vitro-and-in-vivo
#8
Susanne Przybylski, Michaela Gasch, Anne Marschner, Marcus Ebert, Alexander Ewe, Gisa Helmig, Nadja Hilger, Stephan Fricke, Susanne Rudzok, Achim Aigner, Jana Burkhardt
INTRODUCTION: One of the main obstacles in the widespread application of gene therapeutic approaches is the necessity for efficient and safe transfection methods. For the introduction of small oligonucleotide gene therapeutics into a target cell, nanoparticle-based methods have been shown to be highly effective and safe. While immune cells are a most interesting target for gene therapy, transfection might influence basic immune functions such as cytokine expression and proliferation, and thus positively or negatively affect therapeutic intervention...
2017: PloS One
https://www.readbyqxmd.com/read/28463838/engineering-the-rapid-adenovirus-production-and-amplification-rapa-cell-line-to-expedite-the-generation-of-recombinant-adenoviruses
#9
Qiang Wei, Jiaming Fan, Junyi Liao, Yulong Zou, Dongzhe Song, Jianxiang Liu, Jing Cui, Feng Liu, Chao Ma, Xue Hu, Li Li, Yichun Yu, Xiangyang Qu, Liqun Chen, Xinyi Yu, Zhicai Zhang, Chen Zhao, Zongyue Zeng, Ruyi Zhang, Shujuan Yan, Xingye Wu, Yi Shu, Russell R Reid, Michael J Lee, Jennifer Moritis Wolf, Tong-Chuan He
BACKGROUND/AIMS: While recombinant adenoviruses are among the most widely-used gene delivery vectors and usually propagated in HEK-293 cells, generating recombinant adenoviruses remains time-consuming and labor-intense. We sought to develop a rapid adenovirus production and amplification (RAPA) line by assessing human Ad5 genes (E1A, E1B19K/55K, pTP, DBP, and DNA Pol) and OCT1 for their contributions to adenovirus production. METHODS: Stable transgene expression in 293T cells was accomplished by using piggyBac system...
May 3, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28459804/transplantation-of-hgf-gene-engineered-skeletal-myoblasts-improve-infarction-recovery-in-a-rat-myocardial-ischemia-model
#10
Shu-Ling Rong, Xiao-Lin Wang, Cui-Ying Zhang, Zhuo-Hui Song, Lu-Hua Cui, Xiao-Feng He, Xu-Jiong Li, Hui-Jin Du, Bao Li
BACKGROUND: Skeletal myoblast transplantation seems a promising approach for the repair of myocardial infarction (MI). However, the low engraftment efficacy and impaired angiogenic ability limit the clinical efficiency of the myoblasts. Gene engineering with angiogenic growth factors promotes angiogenesis and enhances engraftment of transplanted skeletal myoblasts, leading to improved infarction recovery in myocardial ischemia. The present study evaluated the therapeutic effects of hepatocyte growth factor (HGF) gene-engineered skeletal myoblasts on tissue regeneration and restoration of heart function in a rat MI model...
2017: PloS One
https://www.readbyqxmd.com/read/28454458/survivin-downregulation-using-sirna-nanoliposomes-inhibits-cell-proliferation-and-promotes-the-apoptosis-of-mhcc-97h-hepatic-cancer-cells-an-in-vitro-and-in-vivo-study
#11
Ziqin Liu, Tianyou Wang, Zhaoxia Zhang, Suoqin Tang, Shunqiao Feng, Mei Yue, Mengze Hu, Litian Xuan, Yanfei Chen
At present, survivin is one of the most cancer-specific proteins that has been identified. The present study aimed to investigate the antitumor effects of novel survivin small interfering RNA (siRNA) nanoliposomes targeting survivin in human hepatocellular carcinoma MHCC-97H cells and xenograft mouse models. Survivin-targeted siRNA nanoliposomes were prepared and transfected into MHCC-97H cells and MHCC-97H-bearing nude mice. Survivin expression was analyzed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting...
April 2017: Oncology Letters
https://www.readbyqxmd.com/read/28454422/the-role-of-mrp1-in-the-multidrug-resistance-of-colorectal-cancer
#12
Dongxing Cao, Shaolan Qin, Yifei Mu, Ming Zhong
The role of multidrug resistance associated protein 1 (MRP1) in the multidrug resistance (MDR) of colorectal cancer (CRC) remains unclear. The present study aimed to investigate the effect of MRP1 in MDR CRC and its therapeutic potential for the treatment of patients with this disease. The human MDR CRC cell lines HCT-8 and Colo205 were established through stable exposure to 5-florouracil (5-FU) over a 5-month period. MRP1 was knocked-down in MDR CRC cells through the transfection of short hairpin RNA targeting MRP1 (shMRP1)...
April 2017: Oncology Letters
https://www.readbyqxmd.com/read/28454339/puma-decreases-the-growth-of-prostate-cancer-pc-3-cells-independent-of-p53
#13
Zhengfei Shan, Qingzuo Liu, Yuling Li, Jitao Wu, Dekang Sun, Zhenli Gao
PUMA (p53 upregulated modulator of apoptosis), a member of the B-cell lymphoma 2 (Bcl-2) protein family, is a pro-apoptotic protein. PUMA expression is modulated by the tumor suppressor p53. PUMA has a role in rapid cell death via p53-dependent and -independent mechanisms. To evaluate whether p53 is required for PUMA-mediated apoptosis in prostate cancer cells, p53 protein was silenced in human prostate cancer PC-3 cells by using p53 small interfering RNA (siRNA). The interference efficiency of p53 on RNA and protein levels was detected by reverse transcription-quantitative polymerase chain reaction and western blotting...
March 2017: Oncology Letters
https://www.readbyqxmd.com/read/28453254/gene-transfection-mediated-by-catiomers-requires-free-highly-charged-polymer-chains-to-overcome-intracellular-barriers
#14
Lindomar J C Albuquerque, Carlos E de Castro, Karin A Riske, Maria C Carlan da Silva, Paulo I R Muraro, Vanessa Schmidt, Cristiano Giacomelli, Fernando C Giacomelli
The prospective use of the block copolymers poly(ethylene oxide)113-b-poly[2-(diethylamino)ethyl methacrylate]50 (PEO113-b-PDEA50) and poly[oligo(ethylene glycol)methyl ether methacrylate]70-b-poly[oligo(ethylene glycol)methyl ether methacrylate10-co-2-(diethylamino)ethyl methacrylate47-co-2-(diisopropylamino)ethyl methacrylate47] (POEGMA70-b-P(OEGMA10-co-DEA47-co-DPA47)) as nonviral gene vectors was evaluated. The polymers are able to properly condense DNA into nanosized particles (RH ≈ 75 nm), which are marginally cytotoxic and can be uptaken by cells...
May 5, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28450931/cell-penetrating-peptide-of-sodium-iodide-symporter-effect-on-the-i-131-radiotherapy-on-thyroid-cancer
#15
Yi-Xiang Fan, Zhi-Xin Liang, Qing-Zhu Liu, Han Xiao, Ke-Bin Li, Ji-Zhen Wu
The aim of the present study was to clarify whether the cell penetrating peptide of sodium-iodide symporter (NIS) has an effect on the I-131 radiotherapy of thyroid cancer. Firstly, we combined the HIV-1 TAT peptide (a cell penetrating peptide, dTAT) and established a nanoparticle vector (dTAT NP) to study the delivery efficiency of this cell-penetrating strategy for tumor-targeted gene delivery. dTAT NP was transfected into cultured TPC-1 cells as a model to study the effects of I-131 radiotherapy on thyroid cancer...
March 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28448055/transient-expression-and-cellular-localization-of-recombinant-proteins-in-cultured-insect-cells
#16
Jeffrey A Fabrick, J Joe Hull
Heterologous protein expression systems are used for the production of recombinant proteins, the interpretation of cellular trafficking/localization, and the determination of the biochemical function of proteins at the sub-organismal level. Although baculovirus expression systems are increasingly used for protein production in numerous biotechnological, pharmaceutical, and industrial applications, nonlytic systems that do not involve viral infection have clear benefits but are often overlooked and underutilized...
April 20, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28445602/identification-of-a-novel-mirna-that-increases-transient-protein-expression-in-combination-with-valproic-acid
#17
Hermann-Josef Meyer, Dorothea Reilly, Scott E Martin, Athena W Wong
Transient gene expression in mammalian cells is an efficient process to produce recombinant proteins for various research applications and large molecule therapeutics development. For the first time, we report a screen to identify human microRNAs (miRNAs) that increase titers after polyethylenimine (PEI) mediated transient transfection of a HEK293 cell line. From a library of 875 miRNAs, we identified 2 miRNAs, miR-26a-5p and miR-337-5p, that increased human IgG1 (huIgG1) yields by 50% and 25%, respectively...
April 26, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28442415/retro-inverso-d-peptide-modified-ha-rhb-pdna-core-shell-ternary-nanoparticles-for-the-dual-targeted-delivery-of-short-hairpin-rna-encoding-plasmids
#18
Jijin Gu, Xinyi Chen, Xiaoling Fang, Xianyi Sha
The active targeting of gene carriers is a powerful strategy for improving tumour-specific delivery and therapy. Although numerous L-peptide ligands play significant roles in the active targeting of nanomedicine, retro-inverso D-peptides have been explored as targeting ligands due to their superior stability and bioactivity in vivo. In this study, retro-inverso D-peptide (RIF7)-modified hyaluronic acid (HA)/bioreducible hyperbranched poly(amido amine) (RHB)/plasmid DNA (pDNA) ternary nanoparticles were successfully developed using the layer-by-layer method for the CD44-positive tumour-specific delivery of short hairpin RNA-encoding pDNA through the synergistic reaction of the Anxa1 (tumour vasculature) and CD44 (tumour cell-surface) receptors, which mediated the dual targeting...
April 22, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28440948/novel-reduction-sensitive-micellar-nanoparticles-assembled-from-rituximab-doxorubicin-conjugates-as-smart-and-intuitive-drug-delivery-systems-for-the-treatment-of-non-hodgkin-s-lymphoma
#19
Huabin Yin, Tong Meng, Ling Shu, Min Mao, Lei Zhou, Haiyan Chen, Dianwen Song
In this study, a novel reduction-sensitive drug delivery system, the Rituximab-Doxorubicin (RTX-DOX) micellar nanoparticle (RDMN), was specially designed for targeted delivery and release of DOX in Non-Hodgkin's Lymphoma (NHL) cells. The RDMN was fabricated by self-assembling of amphiphilic RTX-DOX conjugates (RDCs), which were synthesized by conjugating the hydrophilic Fab fragments of RTX (an anti-CD20 monoclonal antibody) and hydrophobic DOXs by a reduction-responsive linker, 3-(2-Pyridyldithio) propionyl hydrazide (PDPH)...
April 25, 2017: Chemical Biology & Drug Design
https://www.readbyqxmd.com/read/28437437/use-of-short-interfering-rna-delivered-by-cationic-liposomes-to-enable-efficient-down-regulation-of-ptpn22-gene-in-human-t-lymphocytes
#20
Valentina Perri, Marsha Pellegrino, Francesca Ceccacci, Anita Scipioni, Stefania Petrini, Elena Gianchecchi, Anna Lo Russo, Serena De Santis, Giovanna Mancini, Alessandra Fierabracci
Type 1 diabetes and thyroid disease are T cell-dependent autoimmune endocrinopathies. The standard substitutive administration of the deficient hormones does not halt the autoimmune process; therefore, development of immunotherapies aiming to preserve the residual hormonal cells, is of crucial importance. PTPN22 C1858T mutation encoding for the R620W lymphoid tyrosine phosphatase variant, plays a potential pathophysiological role in autoimmunity. The PTPN22 encoded protein Lyp is a negative regulator of T cell antigen receptor signaling; R620W variant, leading to a gain of function with paradoxical reduced T cell activation, may represent a valid therapeutic target...
2017: PloS One
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