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https://www.readbyqxmd.com/read/28732241/generation-of-skeletal-myogenic-progenitors-from-human-pluripotent-stem-cells-using-non-viral-delivery-of-minicircle-dna
#1
Jaemin Kim, Vanessa K P Oliveira, Ami Yamamoto, Rita C R Perlingeiro
Currently, the most efficient and promising approach for generating large numbers of engraftable human skeletal myogenic progenitors from pluripotent stem cells requires the conditional in vitro overexpression of PAX7 using lentiviral vectors. Because a non-integrating approach would be preferable to eliminate or minimize the risk associated with random genomic integration, here we investigate whether transient expression of PAX7 using minicircle DNA would enable the generation of functional pluripotent stem cell-derived myogenic progenitors, equivalent to those generated by lentivirus...
July 12, 2017: Stem Cell Research
https://www.readbyqxmd.com/read/28726952/porphyra-polysaccharide-derived-carbon-dots-for-non-viral-co-delivery-of-different-gene-combinations-and-neuronal-differentiation-of-ectodermal-mesenchymal-stem-cells
#2
Jiaxin Chen, Qiang Wang, Jie Zhou, Wenwen Deng, Qingtong Yu, Xia Cao, Jianping Wang, Fengxia Shao, Yang Li, Ping Ma, Myron Spector, Jiangnan Yu, Ximing Xu
In this study, multifunctional fluorescent carbon dots (CDs) were synthesized using a one-pot hydrothermal carbonization reaction, with the naturally-occurring porphyra polysaccharide (PPS) serving as a single carbon source for the first time and ethylenediamine (Ed) acting as the surface passivation agent. The resulting CDs enjoyed a high quantum yield (56.3%), excitation-dependent fluorescence, small size (<10 nm), spherical shape, uniform distribution, positive surface charge, low cytotoxicity and excellent ability to condense macromolecular plasmid DNA...
July 20, 2017: Nanoscale
https://www.readbyqxmd.com/read/28726909/lysine-based-amino-functionalized-lipids-for-gene-transfection-the-protonation-state-in-monolayers-at-the-air-liquid-interface
#3
Stephanie Tassler, Christian Wölk, Christopher Janich, Bodo Dobner, Gerald Brezesinski
Cationic lipids are considered as non-viral carriers for genetic material used in gene therapy. They have no carcinogenic potential and cause low immune response compared to existing viral systems. The protonation degree of these cationic lipids is a crucial parameter for the binding behavior of polynucleotides (e.g., DNA). Newly synthesized peptide-mimic lysine-based amino-functionalized lipids have been investigated in 2D models as monolayers at the air-liquid interface. Standard surface pressure - area isotherms have been measured to prove the layer stability...
July 20, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28722044/tripartite-polyionic-complex-pic-micelles-as-non-viral-vectors-for-mesenchymal-stem-cell-sirna-transfection
#4
Sophie Raisin, Marie Morille, Claire Bony, Danièle Noël, Jean-Marie Devoisselle, Emmanuel Belamie
In the context of regenerative medicine, the use of RNA interference mechanisms has already proven its efficiency in targeting specific gene expression with the aim of enhancing, accelerating or, more generally, directing stem cell differentiation. However, achievement of good transfection levels requires the use of a gene vector. For in vivo applications, synthetic vectors are an interesting option to avoid possible issues associated with viral vectors (safety, production costs, etc.). Herein, we report on the design of tripartite polyionic complex micelles as original non-viral polymeric vectors suited for mesenchymal stem cell transfection with siRNA...
July 19, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28713896/an-optimized-gene-transfection-system-in-weri-rb1-cells
#5
Ying Liu, Zhigang Fan, Kang Li, Fei Deng, Yunfan Xiong, Meixin Liang, Jian Ge
The pathogenesis of Rb1 gene inactivation indicates that gene therapy could be a promising treatment for retinoblastoma. An appropriate gene transfer system is the basis for successful gene therapy; however, little attention has been given to an effective gene transfer system for retinoblastoma therapy in previous studies. This study was designed to provide an optimized transgene system for WERI‑Rb1 cells (W-RBCs). Green fluorescent protein (GFP) was adopted as a reporter. Four classic viral vectors based on retroviruses, recombinant adeno-associated viruses (rAAV2, rAAV2/1), lentiviruses (LVs) and a novel non-viral vector X-treme HP reagent were adopted for W-RBC gene transfection...
July 6, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/28710005/cationic-microbubbles-and-antibiotic-free-miniplasmid-for-sustained-ultrasound-mediated-transgene-expression-in-liver
#6
Simona Manta, Gilles Renault, Anthony Delalande, Olivier Couture, Isabelle Lagoutte, Johanne Seguin, Franck Lager, Pascal Houzé, Patrick Midoux, Michel Bessodes, Daniel Scherman, Michel-Francis Bureau, Corinne Marie, Chantal Pichon, Nathalie Mignet
Despite the increasing number of clinical trials in gene therapy, no ideal methods still allow non-viral gene transfer in deep tissues such as the liver. We were interested in ultrasound (US)-mediated gene delivery to provide long term liver expression. For this purpose, new positively charged microbubbles were designed and complexed with pFAR4, a highly efficient small length miniplasmid DNA devoid of antibiotic resistance sequence. Sonoporation parameters, such as insonation time, acoustic pressure and duration of plasmid injection were controlled under ultrasound imaging guidance...
July 11, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28699870/microrna-mir-214-inhibits-snakehead-vesiculovirus-replication-by-targeting-the-coding-regions-of-viral-n-and-p
#7
Chi Zhang, Lizhu Yi, Shuangshuang Feng, Xueqin Liu, Jianguo Su, Li Lin, Jiagang Tu
Snakeheadvesiculovirus (SHVV), a new member of the family Rhabdoviridae, has caused enormous economic losses in snakehead fish culture during the past years in China; however, little is known about the molecular mechanisms of its pathogenicity. MicroRNAs (miRNAs) are small non-coding RNAs that play important roles in virus infection. In this study, we identified that SHVV infection downregulated miR-214 in striped snakehead (SSN-1) cells in a time- and dose-dependent manner. Notably, transfecting SSN-1 cells with miR-214 mimic significantly inhibitedSHVV replication, whereas miR-214 inhibitor promoted it, suggesting that miR-214 acted as a negative regulator of SHVV replication...
July 12, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28672000/synthesis-and-evaluation-of-l-arabinose-based-cationic-glycolipids-as-effective-vectors-for-pdna-and-sirna-in-vitro
#8
Bo Li, Wanrong Guo, Fan Zhang, Meiyan Liu, Shang Wang, Zhonghua Liu, Shuanglin Xiang, Youlin Zeng
Glycolipids might become a new type of promising non-viral gene delivery systems because of their low cytotoxicity, structural diversity, controllable aqua- and lipo-solubility, appropriate density and distribution of positive charges, high transfer efficiency and potential targeting function. In this study, four kinds of L-arabinose-based cationic glycolipids (Ara-DiC12MA, Ara-DiC14MA, Ara-DiC16MA and Ara-DiC18MA) containing quaternary ammonium as hydrophilic headgroup and two alkane chains as hydrophobic domain were synthesized and characterized...
2017: PloS One
https://www.readbyqxmd.com/read/28667693/in-vitro-and-in-vivo-gene-delivery-using-chitosan-hyaluronic-acid-nanoparticles-influences-of-molecular-mass-of-hyaluronic-acid-and-lyophilization-on-the-transfection-efficiency
#9
Toshinori Sato, Mitsuhiro Nakata, Zhihong Yang, Yu Torizuka, Satoko Kishimoto, Masayuki Ishihara
BACKGROUND: Lyophilization is an effective method for preserving non-viral gene vectors. To improve the stability and transgene expression of lyophilized plasmid DNA (pDNA) complexes, we coated the surfaces of pDNA/chitosan complexes with hyaluronic acid (HA) having varying molecular masses. The transgene expression of pDNA/chitosan/HA ternary complexes was characterized in vitro and in vivo. METHODS: pDNA complexes were lyophilized overnight and the resultant products with spongy, porous consistencies were stored at -30 °C, 4 °C, or 25 °C for two weeks...
July 1, 2017: Journal of Gene Medicine
https://www.readbyqxmd.com/read/28659483/human-bocavirus-1-is-a-novel-helper-for-adeno-associated-virus-replication
#10
Zekun Wang, Xuefeng Deng, Wei Zou, John F Engelhardt, Ziying Yan, Jianming Qiu
Human bocavirus 1 (HBoV1) is an autonomous parvovirus that infects well-differentiated primary human airway epithelia (HAE) in vitro. In human embryonic kidney (HEK) 293 cells, transfection of a duplex HBoV1 genome initiates viral DNA replication and produces progeny virions that are infectious in HAE. HBoV1 takes advantage of signaling pathways in the DNA damage response for efficient genome amplification in both well-differentiated (non-dividing) HAE and dividing HEK293 cells. On the other hand, adeno-associated virus 2 (AAV2) is a helper-dependent dependoparvovirus, and productive AAV2 replication requires co-infection with a helper virus (e...
June 28, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28653538/cationic-polypeptoids-with-optimized-molecular-characteristics-toward-efficient-nonviral-gene-delivery
#11
Lipeng Zhu, Jessica M Simpson, Xin Xu, Hua He, Donghui Zhang, Lichen Yin
The rational design of gene vectors relies on the understanding of their structure-property relationship. Polypeptoids, which are structural isomers of natural polypeptides, hold great potential as gene delivery vectors due to their facile preparation, structural tunability, and most importantly, their desirable proteolytic stability. We herein designed a library of polypeptoids with different cationic side-chain terminal groups, degree of polymerizations (DPs), side-chain lengths, and incorporated aliphatic side chains, to unravel the structure-property relationships so that gene delivery efficiency can be maximized and cytotoxicity can be minimized...
July 19, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28632911/polymer-functionalized-gold-nanoparticles-as-non-viral-gene-delivery-reagents
#12
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
#13
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
#14
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/28624218/targeted-multifunctional-lipid-eco-plasmid-dna-nanoparticles-as-efficient-non-viral-gene-therapy-for-leber-s-congenital-amaurosis
#15
Da Sun, Bhubanananda Sahu, Songqi Gao, Rebecca M Schur, Amita M Vaidya, Akiko Maeda, Krzysztof Palczewski, Zheng-Rong Lu
Development of a gene delivery system with high efficiency and a good safety profile is essential for successful gene therapy. Here we developed a targeted non-viral delivery system using a multifunctional lipid ECO for treating Leber's congenital amaurosis type 2 (LCA2) and tested this in a mouse model. ECO formed stable nanoparticles with plasmid DNA (pDNA) at a low amine to phosphate (N/P) ratio and mediated high gene transfection efficiency in ARPE-19 cells because of their intrinsic properties of pH-sensitive amphiphilic endosomal escape and reductive cytosolic release (PERC)...
June 16, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28618452/murine-melanoma-cells-incomplete-reprogramming-using-non-viral-vector
#16
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/28603974/evolution-of-new-bolaliposomes-using-novel-%C3%AE-tocopheryl-succinate-based-cationic-lipid-and-1-12-disubstituted-dodecane-based-bolaamphiphile-for-efficient-gene-delivery
#17
Mallikarjun Gosangi, Hithavani Rapaka, Venkatesh Ravula, Srilakshmi V Patri
Nonviral 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 require further improvement. Thus, herein, we report a novel lipid formulation by the mixing of two distinct cationic surfactants such as tocopheryl succinate based cationic lipid and 1,12 dodecane based bolaamphiphile and prove it to be a good transfection reagent with its competing potential with the "golden standard", Lipofectamine 3000 (L3K)...
June 27, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28602136/in-situ-forming-hydrogels-with-long-lasting-mir-21-enhances-the-therapeutic-potential-of-msc-by-sustaining-stimulation-of-target-gene
#18
Xianwei Zhang, Yufa Sun, Jianfeng Liu, Zhiyong Yi, Feng Gao, Qiaowei Liu, Yundai Chen
Regulating stem cells by microRNA (miRNA) is promising in regenerative medicine. However, non-viral transfection is usually transient while stably lentiviral transfection is accompanied with oncogenic risk. In the study, we explored the feasibility to retain the microRNAs within biopolymer hydrogels for their long-lasting working and sustaining stimulation of target gene. miRNA-21 (MiR-21), a reported microRNA enhancing the therapeutic potential of mesenchymal stem cells (MSCs) was used. We demonstrated that miR-21 could be efficiently retained within collagen hydrogel after forming complex with cationic polymer polyethylenimine (PEI)...
June 10, 2017: Journal of Biomaterials Science. Polymer Edition
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
#19
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
#20
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
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