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https://www.readbyqxmd.com/read/28216629/matrix-attachment-region-combinations-increase-transgene-expression-in-transfected-chinese-hamster-ovary-cells
#1
Chun-Peng Zhao, Xiao Guo, Si-Jia Chen, Chang-Zheng Li, Yun Yang, Jun-He Zhang, Shao-Nan Chen, Yan-Long Jia, Tian-Yun Wang
Matrix attachment regions (MARs) are cis-acting DNA elements that can increase transgene expression levels in a CHO cell expression system. To investigate the effects of MAR combinations on transgene expression and the underlying regulatory mechanisms, we generated constructs in which the enhanced green fluorescent protein (eGFP) gene flanked by different combinations of human β-interferon and β-globin MAR (iMAR and gMAR, respectively), which was driven by the cytomegalovirus (CMV) or simian virus (SV) 40 promoter...
February 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28210900/false-responses-of-renilla-luciferase-reporter-control-to-nuclear-receptor-tr4
#2
Dongyun Zhang, Sam S Atlasi, Krishna K Patel, Zihao Zhuang, Anthony P Heaney
Renilla luciferase reporter is a widely used internal control in dual luciferase reporter assay system, where its transcription is driven by a constitutively active promoter. However, the authenticity of the Renilla luciferase response in some experimental settings has recently been questioned. Testicular receptor 4 (TR4, also known as NR2C2) belongs to the subfamily 2 of nuclear receptors. TR4 binds to a direct repeat regulatory element in the promoter of a variety of target genes and plays a key role in tumorigenesis, lipoprotein regulation, and central nervous system development...
February 16, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28206953/zinc-finger-protein-zfp36l1-promotes-osteoblastic-differentiation-but-represses-adipogenic-differentiation-of-mouse-multipotent-cells
#3
Kuo-Yun Tseng, Yi-Hsuan Chen, Shankung Lin
Zinc finger protein 36, C3H type-like 1 (ZFP36L1) is a member of the tristetraprolin (TTP) family and its role in the aging-related bone loss is currently unknown. We present evidence that ZFP36L1 expression in rat femurs and bone marrow mesenchymal stem cells (bmMSCs) was down-regulated with aging. ZFP36L1 knockdown decreased osteoblastic differentiation of MC3T3-E1 and C3H10T1/2 cells, and increased adipogenic differentiation of 3T3-L1 and C3H10T1/2 cells, whereas ZFP36L1 overexpression did the opposite. The finding that ZFP36L1 overexpression enhanced osteoblastic and repressed adipogenic differentiation was also corroborated by ex vivo experiments...
February 9, 2017: Oncotarget
https://www.readbyqxmd.com/read/28099942/bigh3-silencing-increases-retinoblastoma-tumor-growth-in-the-murine-sv40-tag-rb-model
#4
Nathalie Allaman-Pillet, Anne Oberson, Daniel F Schorderet
BIGH3, a secreted protein of the extracellular matrix interacts with collagen and integrins on the cell surface. BIGH3 can have opposing functions in cancer, acting either as tumor suppressor or promoter by enhancing tumor progression and angiogenesis. In the eye, BIGH3 is expressed in the cornea and the retinal pigment epithelium and could impact on the development of retinoblastoma, the most common paediatric intraocular neoplasm. Retinoblastoma initiation requires the inactivation of both alleles of the RB1 tumor suppressor gene in the developing retina and tumor progression involves additional genomic changes...
January 14, 2017: Oncotarget
https://www.readbyqxmd.com/read/28052271/establishment-and-characterization-of-an-sv40-large-t-antigen-transduced-porcine-colonic-epithelial-cell-line
#5
Bastian Kaiser, Martina Böttner, Thilo Wedel, Ronald M Brunner, Tom Goldammer, Szilvia Lesko, Gotthold Gäbel, Alexander Gleich, Helga Pfannkuche
Continuous cell lines have become indispensable tools that have enabled investigations into cellular mechanisms by increasing experimental reproducibility and sample availability, and decreasing the use of experimental animals. To facilitate studies of epithelial barrier function of the porcine colon, we aimed to establish an epithelial cell line with an extended replicative capacity. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen (SV40 TAg)...
January 5, 2017: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/28012275/emc1-dependent-stabilization-drives-membrane-penetration-of-a-partially-destabilized-non-enveloped-virus
#6
Parikshit Bagchi, Takamasa Inoue, Billy Tsai
Destabilization of a non-enveloped virus generates a membrane transport-competent viral particle. Here we probe polyomavirus SV40 endoplasmic reticulum (ER)-to-cytosol membrane transport, a decisive infection step where destabilization initiates this non-enveloped virus for membrane penetration. We find that a member of the ER membrane protein complex (EMC) called EMC1 promotes SV40 ER membrane transport and infection. Surprisingly, EMC1 does so by using its predicted transmembrane residue D961 to bind to and stabilize the membrane-embedded partially destabilized SV40, thereby preventing premature viral disassembly...
December 24, 2016: ELife
https://www.readbyqxmd.com/read/27987076/hop-expression-is-regulated-by-p53-and-ras-and-characteristic-of-a-cancer-gene-signature
#7
Stacey A Mattison, Gregory L Blatch, Adrienne L Edkins
The Hsp70/Hsp90 organising protein (HOP) is a co-chaperone essential for client protein transfer from Hsp70 to Hsp90 within the Hsp90 chaperone machine. Although HOP is upregulated in various cancers, there is limited information from in vitro studies on how HOP expression is regulated in cancer. The main objective of this study was to identify the HOP promoter and investigate its activity in cancerous cells. Bioinformatic analysis of the -2500 to +16 bp region of the HOP gene identified a large CpG island and a range of putative cis-elements...
December 16, 2016: Cell Stress & Chaperones
https://www.readbyqxmd.com/read/27896813/transcriptional-and-post-transcriptional-regulation-of-histone-variant-h2a-z-during-sea-urchin-development
#8
Mihai Hajdu, Jasmine Calle, Andrea Puno, Aminat Haruna, César Arenas-Mena
Histone variant H2A.Z promotes chromatin accessibility at transcriptional regulatory elements and is developmentally regulated in metazoans. We characterize the transcriptional and post-transcriptional regulation of H2A.Z in the purple sea urchin Strongylocentrotus purpuratus. H2A.Z depletion by antisense translation-blocking morpholino oligonucleotides during early development causes developmental collapse, in agreement with its previously demonstrated general role in transcriptional multipotency. During H2A...
December 2016: Development, Growth & Differentiation
https://www.readbyqxmd.com/read/27864420/the-landscape-of-viral-expression-reveals-clinically-relevant-viruses-with-potential-capability-of-promoting-malignancy-in-lower-grade-glioma
#9
Zheng Wang, Yajing Hao, Chuan-Ba Zhang, Zhiliang Wang, Xing Liu, Guanzhang Li, Lihua Sun, Jingshan Liang, Jianjun Luo, Dabiao Zhou, Run-Sheng Chen, Tao Jiang
Purpose RNA sequencing (RNA-seq) has recently proved to be effective for revealing novel virus-tumor associations. To get a thorough investigation of virus-glioma associations, we screened viruses in gliomas with RNA-seq data from Chinese Glioma Genome Atlas project (CGGA). Experimental Design In total, 325 samples were enrolled into this study. Reads that failed to map to human genome were aligned to viral genomes and screened for potential virus-derived transcripts. For quantification, VPKM was calculated according to mapped reads weighted by genome sizes and sequencing depth...
November 18, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/27787944/generation-of-immortalised-equine-chondrocytes-with-inducible-sox9-expression-allows-control-of-hypertrophic-differentiation
#10
Saliya Gurusinghe, Bryan Hilbert, Gareth Trope, Lexin Wang, Nadeeka Bandara, Padraig Strappe
Immortalisation of chondrocytes enables long term in vitro culture; however, the chondrogenic capacity of transformed cells varies, thus highlighting the need to develop a proliferative and tuneable chondrocyte cell line where hypertrophic differentiation can be controlled. In this study the SV40 large T antigen and human telomerase reverse transcriptase were employed to immortalise pooled equine chondrocytes through lentiviral vector mediated transduction either singly or on combination. Transformed chondrocytes proliferated stably over multiple passages, but resulted in significantly lower expression of chondrocyte specific collagen II mRNA (p < 0...
October 27, 2016: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/27630227/the-presumed-polyomavirus-viroporin-vp4-of-simian-virus-40-or-human-bk-polyomavirus-is-not-required-for-viral-progeny-release
#11
Stian Henriksen, Terkel Hansen, Jack-Ansgar Bruun, Christine Hanssen Rinaldo
: The minor capsid protein of human BK polyomavirus (BKPyV), VP2, and its N-terminally truncated form, VP3, are both important for viral entry. The closely related simian virus 40 (SV40) reportedly produces an additional truncated form of VP2/3, denoted VP4, apparently functioning as a viroporin promoting progeny release. The VP4 open reading frame is conserved in some polyomaviruses, including BKPyV. In this study, we investigated the role of VP4 in BKPyV replication. By transfecting viral genomes into primary human renal proximal tubule epithelial cells, we demonstrated that unaltered BKPyV and mutants with start codon substitutions in VP4 (VP2M229I and VP2M229A) abolishing putative VP4 production were released at the same level to supernatants...
November 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27617744/enterovirus-a71-dna-launched-infectious-clone-as-a-robust-reverse-genetic-tool
#12
Chee Wah Tan, Han Kang Tee, Michelle Hui Pheng Lee, I-Ching Sam, Yoke Fun Chan
Enterovirus A71 (EV-A71) causes major outbreaks of hand, foot and mouth disease, and is occasionally associated with neurological complications and death in children. Reverse genetics is widely used in the field of virology for functional study of viral genes. For EV-A71, such tools are limited to clones that are transcriptionally controlled by T7/SP6 bacteriophage promoter. This is often time-consuming and expensive. Here, we describe the development of infectious plasmid DNA-based EV-A71 clones, for which EV-A71 genome expression is under transcriptional control by the CMV-intermediate early promoter and SV40 transcriptional-termination signal...
2016: PloS One
https://www.readbyqxmd.com/read/27572655/promoter-optimisation-of-lentiviral-vectors-for-efficient-insulin-gene-expression-in-canine-mesenchymal-stromal-cells-potential-surrogate-beta-cells
#13
Pratigya Gautam, Robert D Foale, Asha Recino, Jing Zhao, Shu Uin Gan, Maja Wallberg, Roy Calne, Andrew M L Lever
BACKGROUND: The lack of an ideal cell type that can be easily acquired, modified to produce insulin, and re-implanted has been a limitation for ex vivo insulin gene therapy. Canine diabetes is currently treated with human insulin and is a good model for human diabetes. Mesenchymal stromal cells (MSCs) are a promising candidate cell type for gene therapy. Here we optimised insulin production using lentiviral transduced canine MSCs aiming to evaluate their ability for use as surrogate beta cells...
August 30, 2016: Journal of Gene Medicine
https://www.readbyqxmd.com/read/27551315/liver-specific-ng37-overexpression-leads-to-diet-dependent-fatty-liver-disease-accompanied-by-cardiac-dysfunction
#14
Xin Zhou, MengMeng Xu, Liyang Wang, Yulian Mu, Rui Feng, Zhilong Dong, Yuexin Pan, Xunzhang Chen, Yongfeng Liu, Shangen Zheng, Donald D Anthony, Jianjie Ma, Williams B Isaacs, Xuehong Xu
BACKGROUND: Environmental factors are well-known causes of diseases. However, aside from a handful of risk indicators, genes' encoding susceptibility to chronic illnesses and their associated environmental triggers are largely unknown. In this era of increasingly rich diets, such genetic predispositions would be immensely helpful from a public health perspective. The novel transgenic mouse model with liver-specific NG37 overexpression characterized in this article identifies the diet-dependent function of NG37 in the pathogenesis of fatty liver disease and cardiac arrhythmia...
2016: Genes & Nutrition
https://www.readbyqxmd.com/read/27462916/p53-elevation-in-human-cells-halt-sv40-infection-by-inhibiting-t-ag-expression
#15
Nir Drayman, Orly Ben-Nun-Shaul, Veronika Butin-Israeli, Rohit Srivastava, Ariel M Rubinstein, Caroline S Mock, Ela Elyada, Yinon Ben-Neriah, Galit Lahav, Ariella Oppenheim
SV40 large T-antigen (T-ag) has been known for decades to inactivate the tumor suppressor p53 by sequestration and additional mechanisms. Our present study revealed that the struggle between p53 and T-ag begins very early in the infection cycle. We found that p53 is activated early after SV40 infection and defends the host against the infection. Using live cell imaging and single cell analyses we found that p53 dynamics are variable among individual cells, with only a subset of cells activating p53 immediately after SV40 infection...
August 16, 2016: Oncotarget
https://www.readbyqxmd.com/read/27458925/incorporation-of-a-nuclear-localization-signal-in-ph-responsive-lah4-l1-peptide-enhances-transfection-and-nuclear-uptake-of-plasmid-dna
#16
Yingying Xu, Wanling Liang, Yingshan Qiu, Marco Cespi, Giovanni F Palmieri, A James Mason, Jenny K W Lam
The major intracellular barriers associated with DNA delivery using nonviral vectors are inefficient endosomal/lysosomal escape and poor nuclear uptake. LAH4-L1, a pH responsive cationic amphipathic peptide, is an efficient DNA delivery vector that promotes the release of nucleic acid into cytoplasm through endosomal escape. Here we further enhance the DNA transfection efficiency of LAH4-L1 by incorporating nuclear localizing signal (NLS) to promote nuclear importation. Four NLSs were investigated: Simian virus 40 (SV40) large T-antigen derived NLS, nucleoplasmin targeting signal, M9 sequence, and the reverse SV40 derived NLS...
September 6, 2016: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/27421099/smad2-overexpression-enhances-adhesion-of-gingival-epithelial-cells
#17
Shoichi Hongo, Tadashi Yamamoto, Keisuke Yamashiro, Masayuki Shimoe, Kazuya Tomikawa, Yuki Ugawa, Shinsuke Kochi, Hidetaka Ideguchi, Hiroshi Maeda, Shogo Takashiba
OBJECTIVE: Gingival epithelial cells play an important role in preventing the initiation of periodontitis, by their hemidesmosomal adhesion to the tooth root surface. Adhesion requires integrin-extracellular matrix (ECM) interactions that are intricately regulated by transforming growth factor-β (TGF-β) signaling. However, the mechanisms underlying the interplay between adhesion molecules and TGF-β, especially the respective roles of Smad2 and Smad3, remain elusive. In this study, we examined the effects of Smad overexpression on gingival epithelial cell adhesion and expression profiles of integrin and ECM-related genes...
November 2016: Archives of Oral Biology
https://www.readbyqxmd.com/read/27382431/coevolution-of-neoplastic-epithelial-cells-and-multilineage-stroma-via-polyploid-giant-cells-during-immortalization-and-transformation-of-mullerian-epithelial-cells
#18
Shiwu Zhang, Imelda Mercado-Uribe, Anil Sood, Robert C Bast, Jinsong Liu
Stromal cells are generally considered to be derived primarily from the host's normal mesenchymal stromal cells or bone marrow. However, the origins of stromal cells have been quite controversial. To determine the role of polyploidy in tumor development, we examined the fate of normal mullerian epithelial cells during the immortalization and transformation process by tracing the expression of SV40 large T antigen. Here we show that immortalized or HRAS-transformed mullerian epithelial cells contain a subpopulation of polyploid giant cells that grow as multicellular spheroids expressing hematopoietic markers in response to treatment with CoCl2...
March 2016: Genes & Cancer
https://www.readbyqxmd.com/read/27357627/advanced-design-of-dumbbell-shaped-genetic-minimal-vectors-improves-non-coding-and-coding-rna-expression
#19
Xiaoou Jiang, Han Yu, Cui Rong Teo, Genim Siu Xian Tan, Sok Chin Goh, Parasvi Patel, Yiqiang Kevin Chua, Nasirah Banu Sahul Hameed, Antonio Bertoletti, Volker Patzel
Dumbbell-shaped DNA minimal vectors lacking nontherapeutic genes and bacterial sequences are considered a stable, safe alternative to viral, nonviral, and naked plasmid-based gene-transfer systems. We investigated novel molecular features of dumbbell vectors aiming to reduce vector size and to improve the expression of noncoding or coding RNA. We minimized small hairpin RNA (shRNA) or microRNA (miRNA) expressing dumbbell vectors in size down to 130 bp generating the smallest genetic expression vectors reported...
September 2016: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/27347338/regulation-of-insulin-like-growth-factor-1-receptor-signaling-by-microrna-4458-in-the-development-of-lumbar-disc-degeneration
#20
Zuo-Qing Liu, Wen-Qin Fu, Shan Zhao, Xin Zhao
A potential role of Insulin-like growth factor 1 (IGF1) receptor/phosphatidylinositol-3 kinase (PI3k)/Akt signaling in the initiation and progression of Lumbar disc degeneration (LDD) has been recently reported. However, the regulation of IGF1 receptor (IGF1R) at post-transcriptional levels in the development of LDD remains unknown. Here, we studied the effects of microRNA-4458 on the expression of IGF1R. We examined the IGF1R levels and microRNA-4458 (miR-4458) levels in the resected LDD discs, compared with the traumatized, non-LDD discs...
2016: American Journal of Translational Research
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