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https://www.readbyqxmd.com/read/29920228/hepatocyte-derived-lipocalin-2-is-a-potential-serum-biomarker-reflecting-tumor-burden-in-hepatoblastoma
#1
Laura Molina, Danielle Bell, Junyan Tao, Morgan Preziosi, Tirthadipa Pradhan-Sundd, Sucha Singh, Minakshi Poddar, Jianhua Luo, Sarangarajan Ranganathan, Maria Chikina, Satdarshan P Monga
Hepatoblastoma (HB) is the most common pediatric liver malignancy. Previously, we reported co-activation of β-catenin and YAP1 in 80% of HB. Hepatic co-expression of active β-catenin and YAP1 via sleeping beauty transposon/transposase and hydrodynamic tail vein injection (SB-HTVI) led to HB development in mice. Here, we identify lipocalin 2 (LCN2) as a target of β-catenin and YAP1 in HB and show that serum LCN2 values positively correlated with tumor burden. Lcn2 was strongly expressed in HB tumor cells in our mouse model...
June 16, 2018: American Journal of Pathology
https://www.readbyqxmd.com/read/29874846/development-of-a-mouse-model-of-prostate-cancer-using-the-sleeping-beauty-transposon-and-electroporation
#2
Hyun-Ji Choi, Han-Byul Lee, Sunyoung Jung, Hyun-Kyu Park, Woori Jo, Sung-Min Cho, Woo-Jin Kim, Woo-Chan Son
The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors in the prostate. In the present study, electroporation was used to enhance the transfection efficiency of the SB transposon. To generate tumors, three constructs (a c-Myc expression cassette, a HRAS (HRas proto-oncogene, GTPase) expression cassette and a shRNA against p53 ) contained within the SB transposon plasmids were directly injected into the prostate...
June 5, 2018: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/29858090/the-antibiotic-free-pfar4-vector-paired-with-the-sleeping-beauty-transposon-system-mediates-efficient-transgene-delivery-in-human-cells
#3
Marie Pastor, Sandra Johnen, Nina Harmening, Mickäel Quiviger, Julie Pailloux, Martina Kropp, Peter Walter, Zoltán Ivics, Zsuzsanna Izsvák, Gabriele Thumann, Daniel Scherman, Corinne Marie
The anti-angiogenic and neurogenic pigment epithelium-derived factor (PEDF) demonstrated a potency to control choroidal neovascularization in age-related macular degeneration (AMD) patients. The goal of the present study was the development of an efficient and safe technique to integrate, ex vivo, the PEDF gene into retinal pigment epithelial (RPE) cells for later transplantation to the subretinal space of AMD patients to allow continuous PEDF secretion in the vicinity of the affected macula. Because successful gene therapy approaches require efficient gene delivery and stable gene expression, we used the antibiotic-free pFAR4 mini-plasmid vector to deliver the hyperactive Sleeping Beauty transposon system, which mediates transgene integration into the genome of host cells...
June 1, 2018: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/29851496/cloning-a-sleeping-beauty-awaiting-the-kiss
#4
Gábor Vajta
The first 20 years of somatic cell nuclear transfer can hardly be described as a success story. Controversially, many factors leading to the fiasco are not intrinsic features of the technique itself. Misunderstandings and baseless accusations alongside with unsupported fears and administrative barriers hampered cloners to overcome the initial challenging period with obvious difficulties that are common features of a radically new approach. In spite of some promising results of mostly sporadic and small-scale experiments, the future of cloning is still uncertain...
June 2018: Cellular Reprogramming
https://www.readbyqxmd.com/read/29848622/a-collection-of-transgenic-medaka-strains-for-efficient-site-directed-transgenesis-mediated-by-phic31-integrase
#5
Tokiro Ishikawa, Satoshi Ansai, Masato Kinoshita, Kazutoshi Mori
Genetic analysis is facilitated by the efficient production of transgenic strains expressing a DNA of interest as a single copy at a designated chromosomal location. However, technical progress toward this goal in medaka fish ( Oryzias latipes ), a vertebrate model organism, has been slow. It is well known that phiC31 integrase enables efficient site-directed transgenesis by catalyzing the recombination of an attP DNA motif in a host genome with an attB motif in a targeting vector. This system was pioneered in medaka using the Sleeping Beauty transposon system, and the attP site was established at three chromosomal locations...
May 30, 2018: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/29846651/sb-driver-analysis-a-sleeping-beauty-cancer-driver-analysis-framework-for-identifying-and-prioritizing-experimentally-actionable-oncogenes-and-tumor-suppressors
#6
Justin Y Newberg, Michael A Black, Nancy A Jenkins, Neal G Copeland, Karen M Mann, Michael B Mann
Cancer driver prioritization for functional analysis of potential actionable therapeutic targets is a significant challenge. Meta-analyses of mutated genes across different human cancer types for driver prioritization has reaffirmed the role of major players in cancer, including KRAS, TP53 and EGFR, but has had limited success in prioritizing genes with non-recurrent mutations in specific cancer types. Sleeping Beauty (SB) insertional mutagenesis is a powerful experimental gene discovery framework to define driver genes in mouse models of human cancers...
May 26, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29792157/long-term-health-and-germline-transmission-in-transgenic-cattle-following-transposon-mediated-gene-transfer
#7
Soo-Young Yum, Song-Jeon Lee, Sin-Gi Park, In-Gang Shin, Sang-Eun Hahn, Woo-Jae Choi, Hee-Soo Kim, Hyeong-Jong Kim, Seong-Hun Bae, Je-Hyeong Lee, Joo-Yeong Moon, Woo-Sung Lee, Ji-Hyun Lee, Choong-Il Lee, Seong-Jin Kim, Goo Jang
BACKGROUND: Transposon-mediated, non-viral gene delivery is a powerful tool for generating stable cell lines and transgenic animals. However, as multi-copy insertion is the preferred integration pattern, there is the potential for uncontrolled changes in endogenous gene expression and detrimental effects in cells or animals. Our group has previously reported on the generation of several transgenic cattle by using microinjection of the Sleeping Beauty (SB) and PiggyBac (PB) transposons and seeks to explore the long-term effects of this technology on cattle...
May 23, 2018: BMC Genomics
https://www.readbyqxmd.com/read/29790684/the-effect-of-music-on-the-cardiac-activity-of-a-fetus-in-a-cardiotocographic-examination
#8
Grażyna Gebuza, Marta Zaleska, Marzena Kaźmierczak, Estera Mieczkowska, Małgorzata Gierszewska
BACKGROUND: Music therapy as an adjunct to treatment is rarely used in perinatology and obstetrics, despite the proven therapeutic effect. Auditory stimulation through music positively impacts the health of adults and infants, its special role being observed in the development of prematurely born neonates. It is equally interesting how music impacts fetuses. OBJECTIVES: The aim of this study is to assess the parameters of fetuses through cardiotocographic recording in women in the 3rd trimester of pregnancy while listening to Pyotr Tchaikovsky's "Sleeping Beauty" and "Swan Lake...
April 24, 2018: Advances in Clinical and Experimental Medicine: Official Organ Wroclaw Medical University
https://www.readbyqxmd.com/read/29766024/integrating-hdad5-35-vectors-as-a-new-platform-for-hsc-gene-therapy-of-hemoglobinopathies
#9
Chang Li, Nikoletta Psatha, Hongjie Wang, Manvendra Singh, Himanshu Bhusan Samal, Wenli Zhang, Anja Ehrhardt, Zsuzsanna Izsvák, Thalia Papayannopoulou, André Lieber
We generated an integrating, CD46-targeted, helper-dependent adenovirus HDAd5/35++ vector system for hematopoietic stem cell (HSC) gene therapy. The ∼12-kb transgene cassette included a β-globin locus control region (LCR)/promoter driven human γ-globin gene and an elongation factor alpha-1 (EF1α)-mgmtP140K expression cassette, which allows for drug-controlled increase of γ-globin-expressing erythrocytes. We transduced bone marrow lineage-depleted cells from human CD46-transgenic mice and transplanted them into lethally irradiated recipients...
June 15, 2018: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/29735000/value-and-payment-in-sleep-medicine
#10
Emerson M Wickwire, Tilak Verma
Value, like beauty, exists in the eye of the beholder. This article places the value of clinical sleep medicine services in historical context and presents a vision for the value-based sleep of the future. First, the history of value and payment in sleep medicine is reviewed from the early days of the field, to innovative disruption, to the widespread adoption of home sleep apnea testing. Next, the importance of economic perspective is discussed, with emphasis on cost containment and cost-shifting between payers, employers, providers, and patients...
April 30, 2018: Journal of Clinical Sleep Medicine: JCSM: Official Publication of the American Academy of Sleep Medicine
https://www.readbyqxmd.com/read/29727572/functional-dna-delivery-enabled-by-lipid-modified-charge-altering-releasable-transporters-carts
#11
Nancy L Benner, Katherine E Near, Michael H Bachmann, Christopher H Contag, Robert M Waymouth, Paul A Wender
Safe and effective DNA delivery systems are required to enable or enhance clinical strategies and research involving gene therapy and DNA vaccinations. To address this delivery problem, a series of charge-altering releasable transporters (CARTs) with varied lipid content were prepared and evaluated for plasmid DNA (pDNA) delivery into cultured cells. These lipid-modified CART co-oligomers were synthesized in only two steps via sequential organocatalytic ring-opening polymerization of lipid-containing cyclic carbonate monomers and morpholinone monomers...
May 4, 2018: Biomacromolecules
https://www.readbyqxmd.com/read/29704684/improved-molecular-platform-for-the-gene-therapy-of-rare-diseases-by-liver-protein-secretion
#12
REVIEW
Mickael Quiviger, Aristeidis Giannakopoulos, Sebastien Verhenne, Corinne Marie, Eleana F Stavrou, Karen Vanhoorelbeke, Zsuzsanna Izsvák, Simon F De Meyer, Aglaia Athanassiadou, Daniel Scherman
Many rare monogenic diseases are treated by protein replacement therapy, in which the missing protein is repetitively administered to the patient. However, in several cases, the missing protein is required at a high and sustained level, which renders protein therapy far from being adequate. As an alternative, a gene therapy treatment ensuring a sustained effectiveness would be particularly valuable. Liver is an optimal organ for the secretion and systemic distribution of a therapeutic transgene product. Cutting edge non-viral gene therapy tools were tested in order to produce a high and sustained level of therapeutic protein secretion by the liver using the hydrodynamic delivery technique...
April 26, 2018: European Journal of Medical Genetics
https://www.readbyqxmd.com/read/29649238/generation-of-multidrug-resistant-human-tissues-by-overexpression-of-the-abcg2-multidrug-transporter-in-embryonic-stem-cells
#13
Zsuzsa Erdei, Anita Schamberger, György Török, Kornélia Szebényi, György Várady, Tamás I Orbán, László Homolya, Balázs Sarkadi, Ágota Apáti
The ABCG2 multidrug transporter provides resistance against various endo- and xenobiotics, and protects the stem cells against toxins and stress conditions. We have shown earlier that a GFP-tagged version of ABCG2 is fully functional and may be used to follow the expression, localization and function of this transporter in living cells. In the present work we have overexpressed GFP-ABCG2, driven by a constitutive (CAG) promoter, in HUES9 human embryonic stem cells. Stem cell clones were generated to express the wild-type and a substrate-mutant (R482G) GFP-ABCG2 variant, by using the Sleeping Beauty transposon system...
2018: PloS One
https://www.readbyqxmd.com/read/29622795/antitumor-activity-of-cd56-chimeric-antigen-receptor-t-cells-in-neuroblastoma-and-sclc-models
#14
Denise L Crossland, Warren L Denning, Sonny Ang, Simon Olivares, Tiejuan Mi, Kirsten Switzer, Harjeet Singh, Helen Huls, Kate S Gold, Bonnie S Glisson, Laurence J Cooper, John V Heymach
The CD56 antigen (NCAM-1) is highly expressed on several malignancies with neuronal or neuroendocrine differentiation, including small-cell lung cancer and neuroblastoma, tumor types for which new therapeutic options are needed. We hypothesized that CD56-specific chimeric antigen receptor (CAR) T cells could target and eliminate CD56-positive malignancies. Sleeping Beauty transposon-generated CD56R-CAR T cells exhibited αβT-cell receptors, released antitumor cytokines upon co-culture with CD56+ tumor targets, demonstrated a lack of fratricide, and expression of cytolytic function in the presence of CD56+ stimulation...
April 6, 2018: Oncogene
https://www.readbyqxmd.com/read/29622087/generation-of-venus-fluorochrome-expressing-transgenic-handmade-cloned-buffalo-embryos-using-sleeping-beauty-transposon
#15
Dharmendra Kumar, Papori Sharma, Kennady Vijayalakshmy, Naresh L Selokar, Pradeep Kumar, Rasika Rajendran, P S Yadav
The objective of this study was to optimise the electroporation conditions for efficient integration of Venus construct in buffalo fetal fibroblasts using Sleeping Beauty (SB) based transposition and to produce Venus expressing transgenic cloned embryos through handmade cloning (HMC) approach. Primary culture of buffalo fetal fibroblast cells was established and subsequently cultured cells were co-transfected with Venus and helper plasmid at different combinations of electroporation condition. In different combinations of voltage, time and plasmid dose, we observed that 300 V, single pulse for 10 ms in 2 mm cuvette and 1...
April 2018: Tissue & Cell
https://www.readbyqxmd.com/read/29587177/advances-in-functional-genetic-screening-with-transposons-and-crispr-cas9-to-illuminate-cancer-biology
#16
REVIEW
Kathryn A O'Donnell
Large-scale genome sequencing studies have identified a wealth of mutations in human tumors and have dramatically advanced the field of cancer genetics. However, the functional consequences of an altered gene in tumor progression cannot always be inferred from mutation status alone. This underscores the critical need for complementary methods to assign functional significance to mutated genes in cancer. Transposons are mobile genetic elements that serve as powerful tools for insertional mutagenesis. Over the last decade, investigators have employed mouse models with on-demand transposon-mediated mutagenesis to perform unbiased genetic screens to identify clinically relevant genes that participate in the pathogenesis of human cancer...
April 2018: Current Opinion in Genetics & Development
https://www.readbyqxmd.com/read/29575648/spontaneous-development-of-intratumoral-heterogeneity-in-a-transposon-induced-mouse-model-of-glioma
#17
Keisuke Sumiyoshi, Hideto Koso, Sumiko Watanabe
Glioma is the most common form of malignant brain cancer in adults. The Sleeping Beauty (SB) transposon-based glioma mouse model allows for effective in vivo analysis of candidate genes. In the present study, we developed a transposon vector that encodes the triple combination of platelet-derived growth factor subunit A (PDGFA), and shRNAs against Nf1 and Trp53 (shNf1/shp53). Initiation and progression of glioma in the brain were monitored by expression of a fluorescent protein. Transduction of the vector into neural progenitor and stem cells (NPC) in the subventricular zone (SVZ) of the neonatal brain induced proliferation of oligodendrocyte precursor cells, and promoted formation of highly penetrant malignant gliomas within 2-4 months...
March 25, 2018: Cancer Science
https://www.readbyqxmd.com/read/29572319/ossabaw-pigs-with-a-pcsk9-gain-of-function-mutation-develop-accelerated-coronary-atherosclerotic-lesions-a-novel-model-for-preclinical-studies
#18
Fang Yuan, Liang Guo, Kyoung-Ha Park, John R Woollard, Kwon Taek-Geun, Kai Jiang, Tamene Melkamu, Bin Zang, Samantha L Smith, Scott C Fahrenkrug, Frank D Kolodgie, Amir Lerman, Renu Virmani, Lilach O Lerman, Daniel F Carlson
BACKGROUND: Ossabaw pigs are unique miniature swine with genetic predisposition to develop metabolic syndrome and coronary atherosclerosis after extended periods receiving atherogenic diets. We have hypothesized that transgenic Ossabaw swine expressing chimp PCSK9 (proprotein convertase subtilisin-like/kexin type 9) containing the D374Y gain of function would develop familial hypercholesterolemia and coronary artery plaques more rapidly than Landrace swine with the same transgene. METHODS AND RESULTS: Ossabaw and Landrace PCSK9 gain-of-function founders were generated by Sleeping Beauty transposition and cloning...
March 23, 2018: Journal of the American Heart Association
https://www.readbyqxmd.com/read/29562762/efficient-non-viral-t-cell-engineering-by-sleeping-beauty-minicircles-diminishing-dna-toxicity-and-mirnas-silencing-the-endogenous-t-cell-receptors
#19
Julian Clauss, Matthias Obenaus, Csaba Miskey, Zoltán Ivics, Zsuzsanna Izsvák, Wolfgang Uckert, Mario Bunse
Transposon-based vectors have entered clinical trials as an alternative to viral vectors for genetic engineering of T cells. However, transposon vectors require DNA transfection into T cells, which were found to cause adverse effects. T-cell viability was decreased in a dose-dependent manner, and DNA-transfected T cells showed a delayed response upon T-cell receptor (TCR) stimulation with regard to blast formation, proliferation, and surface expression of CD25 and CD28. Gene expression analysis demonstrated a DNA-dependent induction of a type I interferon response and interferon-β upregulation...
May 2018: Human Gene Therapy
https://www.readbyqxmd.com/read/29542085/the-enhancer-trap-in-ciona
#20
Yasunori Sasakura
Enhancer trap is a famous application of transposons. This method is useful for the creation of marker transgenic lines that express a reporter gene in tissue- or organ-specific manner, characterization of enhancers in the genome, finding novel patterns of gene expression, and mutagenesis. In Ciona intestinalis, efficient enhancer traps with Minos and Sleeping beauty transposons have been reported. With the enhancer trap lines, the intronic enhancers regulating the expression of the Musashi gene, the compartment in the digestive tube, the presence of enhancers sensitive to the orientation of the gene that they regulate, and the functions of the Hox1 gene have been revealed...
2018: Advances in Experimental Medicine and Biology
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