keyword
https://read.qxmd.com/read/38526634/the-intracellular-visualization-of-exogenous-dna-in-fluorescence-microscopy
#1
JOURNAL ARTICLE
Christina Greitens, Jean-Christophe Leroux, Michael Burger
In the development of non-viral gene delivery vectors, it is essential to reliably localize and quantify transfected DNA inside the cell. To track DNA, fluorescence microscopy methods are commonly applied. These mostly rely on fluorescently labeled DNA, DNA binding proteins fused to a fluorescent protein, or fluorescence in situ hybridization (FISH). In addition, co-stainings are often used to determine the colocalization of the DNA in specific cellular compartments, such as the endolysosomes or the nucleus...
March 25, 2024: Drug Delivery and Translational Research
https://read.qxmd.com/read/38512685/tracking-tau-in-neurons-how-to-transfect-and-track-exogenous-tau-in-primary-neurons
#2
JOURNAL ARTICLE
Sarah Buchholz, Michael Bell-Simons, Hans Zempel
Primary murine neurons have proved to be an essential tool for the general investigation of neuronal polarity, polarized Tau distribution, and Tau-based neuronal dysfunction in disease paradigms. However, mature primary neurons are notoriously difficult to transfect with non-viral approaches and are very sensitive to cytoskeletal manipulation and imaging. Furthermore, standard non-viral transfection techniques require the use of a supportive glial monolayer or high-density cultures, both of which interfere with microscopy...
2024: Methods in Molecular Biology
https://read.qxmd.com/read/38499991/supercharged-coiled-coil-protein-with-n-terminal-decahistidine-tag-boosts-sirna-complexation-and-delivery-efficiency-of-a-lipoproteoplex
#3
JOURNAL ARTICLE
Jonathan W Sun, Joseph S Thomas, Julia M Monkovic, Halle Gibson, Akash Nagapurkar, Joseph A Frezzo, Priya Katyal, Kamia Punia, Farbod Mahoudinobar, P Douglas Renfrew, Jin Kim Montclare
Short interfering RNA (siRNA) therapeutics have soared in popularity due to their highly selective and potent targeting of faulty genes, providing a non-palliative approach to address diseases. Despite their potential, effective transfection of siRNA into cells requires the assistance of an accompanying vector. Vectors constructed from non-viral materials, while offering safer and non-cytotoxic profiles, often grapple with lackluster loading and delivery efficiencies, necessitating substantial milligram quantities of expensive siRNA to confer the desired downstream effects...
March 18, 2024: Journal of Peptide Science
https://read.qxmd.com/read/38495109/aav8-vector-induced-gliosis-following-neuronal-transgene-expression
#4
JOURNAL ARTICLE
Faye McLeod, Elaine McDermott, Shermin Mak, Darren Walsh, Mark Turnbull, Fiona E N LeBeau, Andrew Jackson, Andrew J Trevelyan, Gavin J Clowry
INTRODUCTION: Expression of light sensitive ion channels by selected neurons has been achieved by viral mediated transduction with gene constructs, but for this to have therapeutic uses, for instance in treating epilepsy, any adverse effects of viral infection on the cerebral cortex needs to be evaluated. Here, we assessed the impact of adeno-associated virus 8 (AAV8) carrying DNA code for a soma targeting light activated chloride channel/FusionRed (FR) construct under the CKIIa promoter...
2024: Frontiers in Neuroscience
https://read.qxmd.com/read/38474444/peptide-self-assembly-facilitating-dna-transfection-and-the-application-in-inhibiting-cancer-cells
#5
JOURNAL ARTICLE
Jingyu Wang, Min Ye, Baokuan Zhu
Non-viral vectors have been developing in gene delivery due to their safety and low immunogenicity. But their transfection effect is usually very low, thus limiting the application. Hence, we designed eight peptides (compounds 1 - 8 ). We compared their performances; compound 8 had the best transfection efficacy and biocompatibility. The transfection effect was similar with that of PEI, a most-widely-employed commercial transfection reagent. Atomic force microscope (AFM) images showed that the compound could self-assemble and the self-assembled peptide might encapsulate DNA...
February 21, 2024: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://read.qxmd.com/read/38468277/streamlined-single-step-non-viral-crispr-cas9-knockout-strategy-enhances-gene-editing-efficiency-in-primary-human-chondrocyte-populations
#6
JOURNAL ARTICLE
Simone Ponta, Angela Bonato, Philipp Neidenbach, Valentino F Bruhin, Alexis Laurent, Lee Ann Applegate, Marcy Zenobi-Wong, Goncalo Barreto
BACKGROUND: CRISPR-Cas9-based genome engineering represents a powerful therapeutic tool for cartilage tissue engineering and for understanding molecular pathways driving cartilage diseases. However, primary chondrocytes are difficult to transfect and rapidly dedifferentiate during monolayer (2D) cell culture, making the lengthy expansion of a single-cell-derived edited clonal population not feasible. For this reason, functional genetics studies focused on cartilage and rheumatic diseases have long been carried out in cellular models that poorly recapitulate the native molecular properties of human cartilaginous tissue (e...
March 11, 2024: Arthritis Research & Therapy
https://read.qxmd.com/read/38467553/synthesis-and-biological-properties-of-polyphenol-containing-linear-and-dendrimeric-cationic-peptides
#7
JOURNAL ARTICLE
Artem A Shatilov, Sergey M Andreev, Anastasiya V Shatilova, Evgeny A Turetskiy, Renata A Kurmasheva, Marina O Babikhina, Larisa V Saprygina, Nadezhda N Shershakova, Dar'ya K Bolyakina, Valeriy V Smirnov, Igor P Shilovsky, Musa R Khaitov
Natural polyphenols are promising compounds for the pharmacological control of oxidative stress in various diseases. However, low bioavailability and rapid metabolism of polyphenols in a form of glycosides or aglycones have stimulated the search for the vehicles that would provide their efficient delivery to the systemic circulation. Conjugation of polyphenols with cationic amphiphilic peptides yields compounds with a strong antioxidant activity and ability to pass through biological barriers. Due to a broad range of biological activities characteristic of polyphenols and peptides, their conjugates can be used in the antioxidant therapy, including the treatment of viral, oncological, and neurodegenerative diseases...
January 2024: Biochemistry. Biokhimii︠a︡
https://read.qxmd.com/read/38464936/analysis-of-cleavage-activity-of-dengue-virus-protease-by-co-transfections
#8
JOURNAL ARTICLE
Lekha Gandhi, Musturi Venkataramana
The genome of the dengue virus codes for a single polypeptide that yields three structural and seven non-structural (NS) proteins upon post-translational modifications. Among them, NS protein-3 (NS3) possesses protease activity, involved in the processing of the self-polypeptide and in the cleavage of host proteins. Identification and analysis of such host proteins as substrates of this protease facilitate the development of specific drugs. In vitro cleavage analysis has been applied, which requires homogeneously purified components...
March 5, 2024: Bio-protocol
https://read.qxmd.com/read/38460643/co-delivery-of-siaeg-1-and-doxorubicin-to-treat-osteosarcoma-via-nanomicelles-for-azide-alkyne-click-conjugation-of-poly-l-lysine-dendrons-onto-zein
#9
JOURNAL ARTICLE
Jiadong Pang, Leilei Huang, Yating Lian, Zhijie Yuan, Fen Wang, Li-Ming Zhang
Astrocyte elevated gene-1 (AEG-1) oncogene is a notorious and evolving target in a variety of human malignancies including osteosarcoma. The RNA interference (RNAi) has been clinically proven to effectively knock down specific genes. To successfully implement RNAi in vivo, protective vectors are required not only to protect unstable siRNAs from degradation, but also to deliver siRNAs to target cells with controlled release. Here, we synthesized a Zein-poly(l-lysine) dendrons non-viral modular system that enables efficient siRNA-targeted AEG-1 gene silencing in osteosarcoma and encapsulation of antitumor drugs for controlled release...
March 7, 2024: International Journal of Biological Macromolecules
https://read.qxmd.com/read/38450566/role-of-foxp3-regulatory-t-cells-in-modulating-immune-responses-to-aav-gene-therapy
#10
JOURNAL ARTICLE
Maite Munoz Melero, Moanaro Biswas
Adeno-associated virus (AAV) gene therapy is making rapid strides due to its wide range of therapeutic applications. However, development of serious immune responses to the capsid antigen or the therapeutic transgene product hinders its full clinical impact. Immune suppressive (IS) drug treatments have been used in various clinical trials to prevent the deleterious effects of cytotoxic T cells to the viral vector or transgene, although there isn't a consensus on the best treatment regimen, dosage, or schedule...
March 7, 2024: Human Gene Therapy
https://read.qxmd.com/read/38449351/nontoxic-biodegradable-hyperbranched-poly-%C3%AE-amino-ester-s-for-efficient-sirna-delivery-and-gene-silencing
#11
JOURNAL ARTICLE
Ying Jie Ooi, Chongquan Huang, Kieran Lau, Sing Yian Chew, Jong Gu Park, Mary B Chan-Park
RNA interference (RNAi)-mediated gene silencing is a promising therapeutic approach to treat various diseases, but safe and efficient delivery remains a major challenge to its clinical application. Non-viral gene vectors, such as poly(β-amino esters) (pBAEs), have emerged as a potential candidate due to their biodegradability, low toxicity profile, ease of synthesis, and high gene transfection efficiency for both DNA and siRNA delivery. However, achieving significant gene silencing using pBAEs often requires a large amount of polymer carrier (with polymer/siRNA weight ratio >100) or high siRNA dose (>100 nM), which might potentially exacerbate toxicity concerns during delivery...
March 6, 2024: ACS Applied Materials & Interfaces
https://read.qxmd.com/read/38445883/non-viral-gene-therapy-for-melanoma-using-lysenin-from-eisenia-foetida
#12
JOURNAL ARTICLE
Min Ren, Ling Yang, Liming He, Jie Wang, Wei Zhao, Chunli Yang, Shuai Yang, Hao Cheng, Meijuan Huang, Maling Gou
Earthworms, long utilized in traditional medicine, serve as a source of inspiration for modern therapeutics. Lysenin, a defensive factor in the coelom fluid of the earthworm Eisenia fetida, has multiple bioactivities. However, the inherent toxicity of Lysenin as a pore-forming protein (PFP) restricts its application in therapy. Here, a gene therapy strategy based on Lysenin for cancer treatment is presented. The formulation consists of polymeric nanoparticles complexed with the plasmid encoding Lysenin. After transfection in vitro, melanoma cells can express Lysenin, resulting in necrosis, autophagy, and immunogenic cell death...
March 6, 2024: Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
https://read.qxmd.com/read/38442045/a-high-throughput-microdroplet-based-single-cell-transfection-method-for-gene-knockout-based-on-the-crispr-cas9-system
#13
JOURNAL ARTICLE
Jiayu Sun, Yang Jiao, Fei Pan, Shuk Han Cheng, Dong Sun
The efficient application of the newly developed gene-editing method CRISPR/Cas9 requires more accurate intracellular gene delivery. Traditional delivery approaches, such as lipotransfection and non-viral delivery methods, must contend with major problems to overcome the drawbacks of low efficiency, high toxicity, and cell-type dependency. The high-throughput microdroplet-based single-cell transfection method presented herein provides an alternative method for delivering genome-editing reagents into single living cells...
March 5, 2024: IEEE Transactions on Nanobioscience
https://read.qxmd.com/read/38441386/nanoplatform-based-in-vivo-gene-delivery-systems-for-cancer-therapy
#14
REVIEW
Rui Luo, Hao Le, Qinjie Wu, Changyang Gong
Gene therapy uses modern molecular biology methods to repair disease-causing genes. As a burgeoning therapeutic, it has been widely applied for cancer therapy. Since 1989, there have been numerous clinical gene therapy cases worldwide. However, a few are successful. The main challenge of clinical gene therapy is the lack of efficient and safe vectors. Although viral vectors show high transfection efficiency, their application is still limited by immune rejection and packaging capacity. Therefore, the development of non-viral vectors is overwhelming...
March 5, 2024: Small
https://read.qxmd.com/read/38433377/establishment-of-cos-bk-cells-persistently-infected-with-archetype-bk-polyomavirus
#15
JOURNAL ARTICLE
Souichi Nukuzuma, Hiroshi Onogi, Tetsuro Suzuki
BK polyomavirus (BKPyV) was the first human polyomavirus to be isolated from an immunosuppressed kidney transplant recipient in 1971. BKPyV reactivation causes BKPyV-associated nephropathy and hemorrhagic cystitis. However, the mechanisms underlying BKPyV replication remain unclear. In the present study, we performed the long-term cultivation of COS-7 cells transfected with archetype KOM-5 DNA, which were designated as COS-BK cells. BKPyV derived from COS-BK cells was characterized by analyzing the amount of the virus based on hemagglutination, viral replication, and the production of viral protein 1 (VP1)...
March 3, 2024: Microbiology and Immunology
https://read.qxmd.com/read/38412450/amphiphilic-multi-targeting-copolymer-micelles-efficiently-deliver-pznf580-to-promote-endothelial-cell-proliferation-and-migration
#16
JOURNAL ARTICLE
Chen Li, Qirong Xu, Xiangyan Meng, Xinghong Duo, Yakai Feng
Cationic copolymers are widely used in gene delivery as a non-viral gene vector, but their applications are limited by low transfection efficiency and high cytotoxicity. In order to enhance the transfection efficiency of copolymer micelles to endothelial cells (HUVECs) and reduce their cytotoxicity, this study synthesized an amphipathic multi-targeted copolymer micelle delivery system PCLMD-PPEGMA-NLS-TAT-REDV (TCMs). Gel test results showed that TCMs showed good pZNF580 binding ability and could effectively load the pZNF580 plasmid...
February 27, 2024: Journal of Materials Chemistry. B, Materials for Biology and Medicine
https://read.qxmd.com/read/38410597/mutation-profile-in-liquid-biopsy-tested-by-next-generation-sequencing-in-mexican-patients-with-non-small-cell-lung-carcinoma-and-its-impact-on-survival
#17
JOURNAL ARTICLE
José Fabián Martínez-Herrera, Gisela Sánchez Domínguez, Juan J Juárez-Vignon Whaley, Sonia Carrasco-Cara Chards, Claudia López Vrátný, Jordi Guzmán Casta, Rodrigo F Riera Sala, Jorge A Alatorre-Alexander, Alec Seidman Sorsby, Mayte Cruz Zermeño, Emilio Conde Flores, Rodrigo R Flores-Mariñelarena, Carla P Sánchez-Ríos, Luis M Martínez-Barrera, Raquel Gerson-Cwilich, Patricio Santillán-Doherty, José C Jiménez López, William López Hernández, Jerónimo R Rodríguez-Cid
BACKGROUND: Lung cancer represents a significant global health concern, often diagnosed in its advanced stages. The advent of massive DNA sequencing has revolutionized the landscape of cancer treatment by enabling the identification of target mutations and the development of tailored therapeutic approaches. Unfortunately, access to DNA sequencing technology remains limited in many developing countries. In this context, we emphasize the critical importance of integrating this advanced technology into healthcare systems in developing nations to improve treatment outcomes...
January 30, 2024: Journal of Thoracic Disease
https://read.qxmd.com/read/38402723/pei-a-new-transfection-method-augments-the-inhibitory-effect-of-rbm5-on-prostate-cancer
#18
JOURNAL ARTICLE
Xijia Zhou, Yingshu Cao, Ranwei Li, Xin Di, Yanqiao Wang, Ke Wang
PEI is a cationic polymer, serving as a non-viral transfection carrier grounded in nanotechnology that enhances transfection efficiency via the proton sponge effect. RBM5 is an RNA-binding protein that can inhibit tumor development. This study involved the transfection of RBM5 in prostate cancer cells with PEI, Lipo2000, and their combination. Transwell and wound healing assays were used to observe invasion and migration of prostate cancer cells and flow cytometry was used to observe the apoptosis. Detect the expression of invasion and migration-related protein MMP9 through western blotting experiment...
February 21, 2024: Biochemical and Biophysical Research Communications
https://read.qxmd.com/read/38401849/cyclization-enhanced-poly-%C3%AE-amino-ester-s-vectors-for-efficient-crispr-gene-editing-therapy
#19
JOURNAL ARTICLE
Xianqing Wang, Yinghao Li, A Sigen, Jing Lyu, Xi Wang, Zhonglei He, Irene Lara-Sáez, Ming Li, Wenxin Wang
Among non-viral gene delivery vectors, poly(β-amino ester)s (PAEs) are one of the most versatile candidates because of their wide monomer availability, high polymer flexibility, and superior gene transfection performance both in vitro and in vivo. Over two decades, PAEs have evolved from linear to highly branched structures, significantly enhancing gene delivery efficacy. Building on the proven efficient sets of monomers in highly branched PAEs (HPAEs), this work introduced a new class of cyclic PAEs (CPAEs) constructed via an A2  + B4  + C2 cyclization synthesis strategy and identified their markedly improved gene transfection capabilities in gene delivery applications...
February 22, 2024: Journal of Controlled Release
https://read.qxmd.com/read/38399344/carbon-based-nanostructures-as-emerging-materials-for-gene-delivery-applications
#20
REVIEW
Sara Yazdani, Mehrdad Mozaffarian, Gholamreza Pazuki, Naghmeh Hadidi, Ilia Villate-Beitia, Jon Zárate, Gustavo Puras, Jose Luis Pedraz
Gene therapeutics are promising for treating diseases at the genetic level, with some already validated for clinical use. Recently, nanostructures have emerged for the targeted delivery of genetic material. Nanomaterials, exhibiting advantageous properties such as a high surface-to-volume ratio, biocompatibility, facile functionalization, substantial loading capacity, and tunable physicochemical characteristics, are recognized as non-viral vectors in gene therapy applications. Despite progress, current non-viral vectors exhibit notably low gene delivery efficiency...
February 18, 2024: Pharmaceutics
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