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sv40 promoter

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...
September 14, 2016: Journal of Virology
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
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
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
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...
July 21, 2016: Oncotarget
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
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...
June 29, 2016: Archives of Oral Biology
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
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
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
Iveta Bartonkova, Aneta Grycova, Zdenek Dvorak
Variety of xenobiotics, including therapeutically used vitamin D analogues or environmental and alimentary endocrine disruptors, may interfere with vitamin D receptor (VDR) signaling, with serious physiological or pathophysiological consequences. Therefore, it is of topical interest to have reliable and efficient in vitro screening tools for the identification of agonists and activators of human VDR. We present here two novel stably transfected human reporter cell lines allowing rapid, high-throughput, and selective identification of VDR agonists and activators...
July 18, 2016: Chemical Research in Toxicology
Marieke A de Graaff, Jamie S E Yu, Hannah C Beird, Davis R Ingram, Theresa Nguyen, Jeffrey Juehui Liu, Svetlana Bolshakov, Károly Szuhai, Pierre Åman, Keila E Torres, Dina Lev, Torsten O Nielsen, Judith V M G Bovée, Alexander J Lazar, Neeta Somaiah
Myxoid liposarcoma has the pathognomonic fusion oncogene FUS-DDIT3 encoding a chimeric transcription factor. Metastatic risk is higher with an increased round cell component and has been linked to aberrations involving the IGFR/PI3K/AKT pathway. These molecular insights have yet to translate to targeted therapies, and the lack of experimental models is a major hindrance. We describe the initial in-depth characterization of a new cell line (DL-221) and establishment of a mouse xenograft model. The cell line DL-221 was derived from a metastatic pleural lesion showing myxoid and round cell histology...
August 2016: Laboratory Investigation; a Journal of Technical Methods and Pathology
Xiao-Yin Wang, Jun-He Zhang, Xi Zhang, Qiu-Li Sun, Chun-Peng Zhao, Tian-Yun Wang
We previously demonstrated that the characteristic sequence of matrix attachment regions (MARs) allows transgenes to be maintained episomally in CHO cells. In the present study, six commonly used promoters from human cytomegalovirus major immediate-early (CMV), simian vacuolating virus 40 (SV40), Rous sarcoma virus, Homo sapiens ubiquitin C, phosphoglycerate kinase, and β-globin, respectively, were evaluated to determine their effects on transgene expression and stability in CHO cells stably transfected via the episomal vector harbouring characteristic MAR motifs...
2016: Scientific Reports
Yingli Shi, Jianhai Xiang, Guangzhou Zhou, Tetsuzan Benny Ron, Hsin-I Tong, Wen Kang, Si Sun, Yuanan Lu
A newly isolated Pacific white shrimp (Litopenaeus vannamei) beta-actin promoter SbaP and its derivative compact construct SbaP (ENX) have recently been demonstrated to promote ectopic gene expression in vitro and in vivo. To further explore the potential transduction application, this newly isolated shrimp promoter SbaP was comparatively tested with cytomegalovirus (CMV), simian virus 40 (SV40), polyhedrin (Polh), and white spot syndrome virus immediate early gene 1 (WSSV ie1) four constitutive promoters and a beta-actin promoter (TbaP) from tilapia fish to characterize its promoting function in eight different cell lines...
June 2016: Marine Biotechnology
Zhengri Li, Lian Cui, Jee Myung Yang, Hyo Seok Lee, Ji Suk Choi, Je Moon Woo, Seul Ki Lim, Kyung Chul Yoon
PURPOSE: To investigate the wound healing effect of adiponectin eye drops following corneal alkali burn. MATERIALS AND METHODS: A chemical burn was induced using 0.1 M NaOH in both adenovirus 12-SV40 hybrid-transformed human corneal epithelial (HCE-2) cells and C57BL/6 mice. The injured HCE-2 and mice were then treated using either 0.1% hyaluronic acid (HA) or adiponectin at 0.0001%, 0.001%, or 0.01% concentration. The viability of the HCE-2 cells was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay...
May 9, 2016: Current Eye Research
Yi-Chao Hsu, Chien-Yu Kao, Yu-Fen Chung, Don-Ching Lee, Jen-Wei Liu, Ing-Ming Chiu
: Fibroblast growth factor 1 (FGF1) binds and activates FGF receptors, thereby regulating cell proliferation and neurogenesis. Human FGF1 gene 1B promoter (-540 to +31)-driven SV40 T antigen has been shown to result in tumorigenesis in the brains of transgenic mice. FGF1B promoter (-540 to +31)-driven green fluorescent protein (F1BGFP) has also been used in isolating neural stem cells (NSCs) with self-renewal and multipotency from developing and adult mouse brains. In this study, we provide six lines of evidence to demonstrate that FGF1/FGFR signaling is implicated in the expression of Aurora A (AurA) and the activation of its kinase domain (Thr288 phosphorylation) in the maintenance of glioblastoma (GBM) cells and NSCs...
April 30, 2016: Experimental Cell Research
Q Lu, T H Ganjawala, E Ivanova, J G Cheng, D Troilo, Z-H Pan
Adeno-associated virus (AAV) vectors have been a powerful gene delivery vehicle to the retina for basic research and gene therapy. For many of these applications, achieving cell type-specific targeting and high transduction efficiency is desired. Recently, there has been increasing interest in AAV-mediated gene targeting to specific retinal bipolar cell types. A 200-bp enhancer in combination with a basal SV40 promoter has been commonly used to target transgenes into ON-type bipolar cells. In the current study, we searched for additional cis-regulatory elements in the mGluR6 gene for improving AAV-mediated transduction efficiency into retinal bipolar cells...
August 2016: Gene Therapy
Katharina Schmidt, Simon Keiser, Viola Günther, Oleg Georgiev, Hans H Hirsch, Walter Schaffner, Tobias Bethge
The non-coding control region (NCCR) of polyomaviruses includes the promoters for early and late genes, a transcription enhancer and the origin of DNA replication. Particularly virulent variants of the human pathogens BKPyV and JCPyV, as well as of simian virus 40 (SV40), occur in vitro and in vivo. These strains often harbour rearrangements in their NCCR, typically deletions of some DNA segment(s) and/or duplications of others. Using an SV40-based model system we provide evidence that duplications of enhancer elements, whether from SV40 itself or from the related BKPyV and JCPyV, increase early gene transcription and replicative capacity...
July 2016: Journal of General Virology
Yong Wu, Tian-Ying Zhang, Lin-Lin Fang, Zi-Xuan Chen, Liu-Wei Song, Jia-Li Cao, Lin Yang, Quan Yuan, Ning-Shao Xia
The stable HBV-replicating cell lines, which carry replication-competent HBV genome stably integrated into the genome of host cell, are widely used to evaluate the effects of antiviral agents. However, current methods to generate HBV-replicating cell lines, which are mostly dependent on random integration of foreign DNA via plasmid transfection, are less-efficient and time-consuming. To address this issue, we constructed an all-in-one Sleeping Beauty transposon system (denoted pTSMP-HBV vector) for robust generation of stable cell lines carrying replication-competent HBV genome of different genotype...
August 2016: Journal of Virological Methods
Hong Wang, Liping Wen, Qingqing Yuan, Min Sun, Minghui Niu, Zuping He
Within the seminiferous tubules there are two major cell types, namely male germ cells and Sertoli cells. Recent studies have demonstrated that male germ cells and Sertoli cells can have significant applications in treating male infertility and other diseases. However, primary male germ cells are hard to proliferate in vitro and the number of spermatogonial stem cells is scarce. Therefore, methods that promote the expansion of these cell populations are essential for their use from the bench to the bed side...
August 2016: Reproduction: the Official Journal of the Society for the Study of Fertility
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