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Non-viral AND transfection

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https://www.readbyqxmd.com/read/28924921/mg-2-induced-dna-compaction-condensation-and-phase-separation-in-gene-delivery-vehicles-based-on-zwitterionic-phospholipids-a-dynamic-light-scattering-and-surface-enhanced-raman-spectroscopic-study
#1
Erhan Süleymanoğlu
Despite the significant efforts towards applying improved non-destructive and label-free measurements of biomolecular structures of lipid-based gene delivery vectors, little is achieved in terms of their structural relevance in gene transfections. Better understanding of structure-activity relationships of lipid-DNA complexes and their gene expression efficiencies thus becomes an essential issue. Raman scattering offers a complimentary measurement technique for following the structural transitions of both DNA and lipid vesicles employed for their transfer...
September 18, 2017: Journal of Biological Inorganic Chemistry: JBIC
https://www.readbyqxmd.com/read/28920716/transposons-moving-forward-from-preclinical-studies-to-clinical-trials
#2
Jaitip Tipanee, Thierry VandenDriessche, Marinee K Chuah
Transposons have emerged as promising vectors for gene therapy that can potentially overcome some of the limitations of commonly used viral vectors. Transposons stably integrate into the target cell genome, enabling persistent expression of therapeutic genes. Transposons have evolved from being used as basic tools in biomedical research to bona fide therapeutics. Currently, the most promising transposons for gene therapy applications are derived from Sleeping Beauty (SB) or piggyBac (PB). Stable transposition requires co-delivery of the transposon DNA with the corresponding transposase gene, mRNA, or protein...
August 22, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28918039/crispr-cas9-mediated-knockin-application-in-cell-therapy-a-non-viral-procedure-for-bystander-treatment-of-glioma-in-mice
#3
Oscar Meca-Cortés, Marta Guerra-Rebollo, Cristina Garrido, Salvador Borrós, Nuria Rubio, Jeronimo Blanco
The use of non-viral procedures, together with CRISPR/Cas9 genome-editing technology, allows the insertion of single-copy therapeutic genes at pre-determined genomic sites, overcoming safety limitations resulting from random gene insertions of viral vectors with potential for genome damage. In this study, we demonstrate that combination of non-viral gene delivery and CRISPR/Cas9-mediated knockin via homology-directed repair can replace the use of viral vectors for the generation of genetically modified therapeutic cells...
September 15, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28917642/efficient-gene-editing-via-non-viral-delivery-of-crispr-cas9-system-using-polymeric-and-hybrid-microcarriers
#4
Alexander S Timin, Albert R Muslimov, Kirill V Lepik, Olga S Epifanovskaya, Alena I Shakirova, Ulrike Mock, Kristoffer Riecken, Maria V Okilova, Vladislav S Sergeev, Boris V Afanasyev, Boris Fehse, Gleb B Sukhorukov
CRISPR-Cas9 is a revolutionary genome-editing technology that has enormous potential for the treatment of genetic diseases. However, the lack of efficient and safe, non-viral delivery systems has hindered its clinical application. Here, we report on the application of polymeric and hybrid microcarriers, made of degradable polymers such as polypeptides and polysaccharides and modified by silica shell, for delivery of all CRISPR-Cas9 components. We found that these microcarriers mediate more efficient transfection than a commercially available liposome-based transfection reagent (>70% vs...
September 13, 2017: Nanomedicine: Nanotechnology, Biology, and Medicine
https://www.readbyqxmd.com/read/28911210/a20-an-essential-component-of-the-ubiquitin-editing-protein-complex-is-a-negative-regulator-of-inflammation-in-human-myometrium-and-foetal-membranes
#5
Martha Lappas
STUDY QUESTION: Does A20 regulate mediators involved in the terminal processes of human labour in primary myometrial and amnion cells? SUMMARY ANSWER: A20 is a nuclear factor-kappa B (NF-κB) responsive gene that acts as a negative regulator of NF-κB-induced expression of pro-labour mediators. WHAT IS KNOWN ALREADY: Inflammation is commonly implicated in spontaneous preterm birth and the processes involved in rupture of foetal membranes and uterine contractions...
September 1, 2017: Molecular Human Reproduction
https://www.readbyqxmd.com/read/28905887/an-efficient-non-viral-dendritic-vector-for-gene-delivery-in-tissue-engineering
#6
D P Walsh, A Heise, F J O'Brien, S-A Cryan
Recent developments within the field of tissue engineering (TE) have shown that biomaterial scaffold systems can be augmented via the incorporation of gene therapeutics. The objective of this study was to assess the potential of the activated polyamidoamine dendrimer (dPAMAM) transfection reagent (Superfect(TM)) as a gene delivery system to mesenchymal stem cells (MSCs) in both monolayer and 3D culture on collagen based scaffolds. dPAMAM-pDNA polyplexes at a mass ratio (M:R) 10:1 (dPAMAM : pDNA) (1 ug pDNA) were capable of facilitating prolonged reporter gene expression in monolayer MSCs which was superior to that facilitated using polyethylenimine (PEI)-pDNA polyplexes (2 ug pDNA)...
September 14, 2017: Gene Therapy
https://www.readbyqxmd.com/read/28899926/peptides-mediating-dna-transport-on-microtubules-and-their-impact-on-non-viral-gene-transfer-efficiency
#7
Patrick Midoux, Lucie Pigeon, Cristine Gonçalves, Chantal Pichon
Synthetic vectors such as cationic polymers and cationic lipids remain attractive tools for non-viral gene transfer which is a complex process whose effectiveness relies on the ability to deliver a plasmid DNA (pDNA) into the nucleus of non-dividing cells. Once in the cytosol, the transport of pDNAs towards the nuclear envelop is strongly impaired by their very low cytosolic mobility due to their large size. To promote their movement towards the cell nucleus, few strategies have been implemented to exploit dynein, the microtubule's motor protein, for propagation of cytosolic pDNA along the microtubules (MTs) towards the cell nucleus...
September 12, 2017: Bioscience Reports
https://www.readbyqxmd.com/read/28899816/enzyme-crosslinked-gene-activated-matrix-for-the-induction-of-mesenchymal-stem-cells-in-osteochondral-tissue-regeneration
#8
Yi-Hsuan Lee, Hsi-Chin Wu, Chia-Wei Yeh, Chen-Hsiang Kuan, Han-Tsung Liao, Horng-Chaung Hsu, Jui-Che Tsai, Jui-Sheng Sun, Tzu-Wei Wang
The development of osteochondral tissue engineering is an important issue for the treatment of traumatic injury or aging associated joint disease. However, the different compositions and mechanical properties of cartilage and subchondral bone show the complexity of this tissue interface, making it challenging for the design and fabrication of osteochondral graft substitute. In this study, a bilayer scaffold is developed to promote the regeneration of osteochondral tissue within a single integrated construct...
September 9, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28887968/polyethyleneimine-grafted-short-halloysite-nanotubes-for-gene-delivery
#9
Zheru Long, Jun Zhang, Yan Shen, Changren Zhou, Mingxian Liu
Inorganic nanoparticles have attracted much attentions in gene delivery because of their desirable characteristics including low toxicity, well-controlled characteristics, high gene delivery efficiency, and multi-functionalities. Here, natural occurred halloysite nanotubes (HNTs) were developed as a novel non-viral gene vector. To increase the efficiency of endocytosis, HNTs were firstly shortened into an appropriate size (~200nm). Then polyethyleneimine (PEI) was grafted onto HNTs to bind green fluorescence protein (GFP) labeled pDNA...
December 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
https://www.readbyqxmd.com/read/28884679/the-integration-of-a-macrophage-adapted-live-vaccine-strain-of-equine-infectious-anaemia-virus-eiav-in-the-horse-genome
#10
Qiang Liu, Xue-Feng Wang, Cheng Du, Yue-Zhi Lin, Jian Ma, Yu-Hong Wang, Jian-Hua Zhou, Xiaojun Wang
Integration is an important feature of retroviruses and retrovirus-based therapeutic transfection vectors. The non-primate lentivirus equine infectious anaemia virus (EIAV) primarily targets macrophages/monocytes in vivo. Investigation of the integration features of EIAVDLV121 strains, which are adapted to donkey monocyte-derived macrophages (MDMs), is of great interest. In this study, we analysed the integration features of EIAVDLV121 in equine MDMs during in vitro infection. Our previously published integration sites (IS) for EIAVFDDV13 in fetal equine dermal (FED) cells were also analysed in parallel as references...
September 7, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28884263/efficient-propagation-of-archetype-jc-polyomavirus-in-cos-7-cells-evaluation-of-rearrangements-within-the-nccr-structural-organization-after-transfection
#11
Carla Prezioso, Daniela Scribano, Anna Bellizzi, Elena Anzivino, Donatella Maria Rodio, Maria Trancassini, Anna Teresa Palamara, Valeria Pietropaolo
John Cunningham virus (JCPyV) is an ubiquitous human pathogen that causes disease in immunocompromised patients. The JCPyV genome is composed of an early region and a late region, which are physically separated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robustness...
September 7, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28877647/improvement-of-in-vivo-expression-of-genes-delivered-by-self-amplifying-rna-using-vaccinia-virus-immune-evasion-proteins
#12
Tim Beissert, Lars Koste, Mario Perkovic, Kerstin C Walzer, Stephanie Erbar, Abderaouf Selmi, Mustafa Diken, Sebastian Kreiter, Özlem Türeci, Ugur Sahin
Among nucleic acid based delivery platforms, self-amplifying RNA (saRNA) vectors are of increasing interest for applications such as transient expression of recombinant proteins and vaccination. saRNA is safe and, due to its capability to amplify intracellularly high protein levels, can by produced from even minute amounts of transfected templates. It is an obstacle to full exploitation of this platform, though, that saRNA induces a strong innate host immune response. In transfected cells pattern recognition receptors sense double-stranded RNA intermediates and via activation of protein kinase R (PKR) and interferon signaling initiate host defense measures including a translational shut-down...
September 6, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28865990/ecm-turnover-stimulated-gene-delivery-through-collagen-mimetic-peptide-plasmid-integration-in-collagen
#13
Morgan A Urello, Kristi L Kiick, Millicent O Sullivan
Gene therapies have great potential in regenerative medicine; however, clinical translation has been inhibited by low stability and limited transfection efficiencies. Herein, we incorporate collagen-mimetic peptide (CMP)-linked polyplexes in collagen scaffolds to increase DNA stability by up to 400% and enable tailorable in vivo transgene expression at 100-fold higher levels and 10-fold longer time periods. These improvements were directly linked to a sustained interaction between collagen and polyplexes that persisted during cellular remodeling, polyplex uptake, and intracellular trafficking...
September 1, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28855426/neural-differentiation-of-fibroblasts-induced-by-intracellular-co-delivery-of-ascl1-brn2-and-foxa1-via-a-non-viral-vector-of-cationic-polysaccharide
#14
Qingtong Yu, Jingjing Chen, Wenwen Deng, Xia Cao, Michael Adu-Frimpong, Jiangnan Yu, Xi-Ming Xu
Direct reprogramming of other somatic cells into neurons is an alternative strategy for the recovery of injured nervous system. In this work we developed a new non-viral gene carrier based on Porphyra yezoensis polysaccharide (PYP). After modification with ethylenediamine, the cationized Porphyra yezoensis polysaccharide (Ed-PYP) was combined with plasmids encoding Ascl1, Brn2 and FoxA1 to form spherical nanoscale particles (Ed-PYP-pABF nanoparticles). Cytotoxicity assay proved that Ed-PYP-pABF nanoparticles had a safer property than Lipofectamine2000 and polyetherimide (PEI)...
August 31, 2017: Biomedical Materials
https://www.readbyqxmd.com/read/28852016/the-histidine-rich-peptide-lah4-l1-strongly-promotes-pamam-mediated-transfection-at-low-nitrogen-to-phosphorus-ratios-in-the-presence-of-serum
#15
Nan Liu, Burkhard Bechinger, Regine Süss
Non-viral vectors are widely used and investigated for the delivery of genetic material into cells. However, gene delivery barriers like lysosomal degradation, serum inhibition and transient gene expression so far still limit their clinical applications. Aiming to overcome these limitations, a pH-sensitive hybrid gene vector (PSL complex) was designed by self-assembly of poly(amidoamine) (PAMAM) dendrimers, the histidine-rich peptide LAH4-L1 and the sleeping beauty transposon system (SB transposon system, a plasmid system capable of efficient and precise genomic insertion)...
August 29, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28842122/malonic-acid-based-cationic-lipids-the-way-to-highly-efficient-dna-carriers
#16
REVIEW
Christian Wölk, Christopher Janich, Udo Bakowsky, Andreas Langner, Gerald Brezesinski
Cationic lipids play an important role as non-viral nucleic acid carriers in gene therapy since 3 decades. This review will introduce malonic acid derived cationic lipids as nucleic acid carriers which appeared in the literature dealing with lipofection 10years ago. The family of amino-functionalized branched fatty acid amides will be presented as well as different generations of malonic acid diamides. Both groups of cationic lipids yield lipid mixtures with highly efficient nucleic acid transfer activities in in-vitro cell culture models...
August 13, 2017: Advances in Colloid and Interface Science
https://www.readbyqxmd.com/read/28836717/transmission-of-penicillium-aurantiogriseum-partiti-like-virus-1-to-a-new-fungal-host-cryphonectria-parasitica-confers-higher-resistance-to-salinity-and-reveals-adaptive-genomic-changes
#17
L Nerva, A Silvestri, M Ciuffo, S Palmano, G C Varese, M Turina
We attempted to transfect six recently characterized virus species to protoplasts of Penicillium janczewskii and Chryphonectria parasitica. None of the recovered P. janczewskii colonies was positive for the transfected viruses, but Penicillium aurantiogriseum partiti-like virus 1 (PaPLV1) was detected in three distinct regenerated C. parasitica colonies. We screened the phenotype of the infected strains in up to 45 different conditions combining different media, salinity and temperatures: our results show that the infected strains grow slower than the virus- free in most of the tested conditions with the exception of halophilic stress in a specific nutrient combination media...
August 24, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28829410/live-imaging-of-primary-cerebral-cortex-cells-using-a-2d-culture-system
#18
Bruna Soares Landeira, Jéssica Alves de Medeiros Araújo, Timm Schroeder, Ulrich Müller, Marcos R Costa
During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or postmitotic neurons. Later, some progenitors switch to a gliogenic fate, adding to the astrocyte and oligodendrocyte populations. Using time-lapse video-microscopy of primary cerebral cortex cell cultures, it is possible to study the cellular and molecular mechanisms controlling the mode of cell division and cell cycle parameters of progenitor cells. Similarly, the fate of postmitotic cells can be examined using cell-specific fluorescent reporter proteins or post-imaging immunocytochemistry...
August 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28826232/minicircle-versus-plasmid-dna-delivery-by-receptor-targeted-polyplexes
#19
Ana Krhac Levacic, Stephan Morys, Susanne Kempter, Ulrich Lächelt, Ernst Wagner
Due to its minimal size and lack of bacterial backbone sequences, minicircle (MC) DNA presents a promising alternative to plasmid DNA (pDNA) for non-viral gene delivery in terms of biosafety and improved gene transfer. Here, luciferase pDNA (pCMV-luc) and analogous MC DNA (MC07.CMV-luc) were formulated into polyplexes with c-Met targeted, PEG-shielded sequence-defined oligoaminoamides, or linear PEI (linPEI) as standard transfection agent. Distinct physicochemical and biological characteristics were observed for polyplexes formed with either pDNA or MC DNA as vectors...
August 21, 2017: Human Gene Therapy
https://www.readbyqxmd.com/read/28796282/disulfide-bond-containing-agamatine-cystaminebisacrylamide-polymer-demonstrates-better-transfection-efficiency-and-lower-cytotoxicity-than-polyethylenimine-in-nih-3t3-cells
#20
Zhen Yang, Yanping Sun, Lei Xian, Zhe Xun, Jiankun Yu, Tianzhi Yang, Xiaoyun Zhao, Cuifang Cai, Dongkai Wang, Pingtian Ding
Previously, we synthesized a non-viral vector containing disulfide bond by polymerization of agamatine (AGM) and N, N'-cystaminebisacrylamide (CBA). In this study, we investigated the transfection efficiency of disulfide bond (-SS-) containing AGM-CBA polymer in gene delivery into NIH/3T3 cells, and examined the factors affecting its transfection efficiency by comparing with polyethylenimine (PEI). In addition, experiments were carried out to determine the mechanisms of cell entry pathways and intracellular behavior of AGM-CBA/pDNA polyplexes...
August 10, 2017: Journal of Cellular Biochemistry
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