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In vivo gene silencing

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https://www.readbyqxmd.com/read/27911980/cell-based-dna-demethylation-detection-system-for-screening-of-epigenetic-drugs-in-2d-3d-and-xenograft-models
#1
Khushboo Agrawal, Viswanath Das, Miroslav Otmar, Marcela Krečmerová, Petr Džubák, Marián Hajdúch
Aberrant DNA methylation that results in silencing of genes has remained a significant interest in cancer research. Despite major advances, the success of epigenetic therapy is elusive due to narrow therapeutic window. A wide variety of naturally occurring epigenetic agents and synthetic molecules that can alter methylation patterns exist, however, their usefulness in epigenetic therapy remains unknown. This underlines the need for effective tumor models for large-scale screening of drug candidates with potent hypomethylation activity...
December 2, 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/27911668/association-of-5-hydroxymethylation-and-5-methylation-of-dna-cytosine-with-tissue-specific-gene-expression
#2
V K Chaithanya Ponnaluri, Kenneth C Ehrlich, Guoqiang Zhang, Michelle Lacey, Douglas Johnston, Sriharsa Pradhan, Melanie Ehrlich
Differentially methylated or hydroxymethylated regions (DMRs) in mammalian DNA are often associated with tissue-specific gene expression but the functional relationships are still being unraveled. To elucidate these relationships, we studied 16 human genes containing myogenic DMRs by analyzing profiles of their epigenetics and transcription and quantitatively assaying 5-hydroxymethylcytosine (5 hmC) and 5-methylcytosine (5 mC) at specific sites in these genes in skeletal muscle (SkM), myoblasts, heart, brain, and diverse other samples...
December 2, 2016: Epigenetics: Official Journal of the DNA Methylation Society
https://www.readbyqxmd.com/read/27911494/determining-functional-aptamer-protein-interaction-by-biolayer-interferometry
#3
Xinhui Lou, Martin Egli, Xianbin Yang
Short single-stranded nucleic acids called aptamers are widely being explored as recognition molecules of high affinity and specificity for binding a wide range of target molecules, particularly protein targets. In biolayer interferometry (BLI), a simple Dip-and-Read approach in which the aptamer-coated biosensors are dipped into microplate wells is used to study the interactions between an aptamer and its target protein. Here we describe the protocol for the analysis of the interaction between a well-characterized anti-thrombin RNA aptamer with thrombin (Basic Protocol)...
December 1, 2016: Current Protocols in Nucleic Acid Chemistry
https://www.readbyqxmd.com/read/27911267/oncogenic-secretory-clusterin-in-hepatocellular-carcinoma-expression-at-early-staging-and-emerging-molecular-target
#4
Wenjie Zheng, Min Yao, Qi Qian, Wenli Sai, Liwei Qiu, Junling Yang, Wei Wu, Zhizhen Dong, Dengfu Yao
Secretory clusterin (sCLU) is associated with hepatocellular carcinoma (HCC) progression by contributing to angiogenesis, chemoresistance, cell survival, and metastasis. However, the sCLU expression at early stage of HCC progression remains to be clarified. In this study, the alteration of sCLU oncogenicity was firstly evaluated in HCC- and their para-cancerous- tissues. The incidence of sCLU expression in HCC was significantly higher than that in their non-tumorous tissues at message RNA (mRNA) or protein level, gradually increasing with tumor-node-metastasis (TNM) staging...
November 29, 2016: Oncotarget
https://www.readbyqxmd.com/read/27908642/negative-regulation-of-nlrp3-inflammasome-by-sirt1-in-vascular-endothelial-cells
#5
Yanxiang Li, Xiaofeng Yang, Yanhao He, Weirong Wang, Jiye Zhang, Wei Zhang, Ting Jing, Bo Wang, Rong Lin
NLRP3 inflammasome not only functions as a critical effector in innate immunity, but also triggers the production of proinflammatory cytokines involved in inflammation-associated diseases. Sirtuin 1 (SIRT1) plays an important role in the regulation of cellular inflammation. However, whether the activation of NLRP3 inflammasome is regulated by SIRT1 remains unknown. In this study, we investigated the regulatory effect of SIRT1 on NLRP3 inflammasome and the underlying mechanisms. We found that lipopolysaccharide (LPS) and adenosine triphosphate (ATP)-induced the activation of NLRP3 inflammasome in human umbilical vein endothelial cells (HUVECs)...
November 4, 2016: Immunobiology
https://www.readbyqxmd.com/read/27908592/activation-of-gpr30-stimulates-gtp-binding-of-g%C3%AE-i1-protein-to-sustain-activation-of-erk1-2-in-inhibition-of-prostate-cancer-cell-growth-and-modulates-metastatic-properties
#6
Lau Kin-Mang, Ma Fanny Man-Ting, Xia Jenny Tian, Chan Queeny Kwan Yi, Ng Chi-Fai, To Ka-Fai
Previously, we reported that GPR30 activation by the receptor-specific, non-estrogenic ligand G-1 inhibited in vitro and in vivo growth of prostate cancer (PCa) cells via sustained Erk1/2 activation. Mechanism underlying the sustained Erk1/2 activation for PCa cell growth inhibition remains unclear. Here we report that G-1, through GPR30, activated Gαi1 proteins to sustain Erk1/2 activation but failed to activate adenylyl cyclase (AC) for cAMP production in PCa cells. The chemical-induced activation of AC-cAMP-PKA signaling attenuated Erk1/2 activity and blocked the cell growth inhibitory effects of G-1...
November 28, 2016: Experimental Cell Research
https://www.readbyqxmd.com/read/27904699/generation-of-induced-pluripotent-stem-cells-with-high-efficiency-from-human-embryonic-renal-cortical-cells
#7
Ling Yao, Ruifang Chen, Pu Wang, Qi Zhang, Hailiang Tang, Huaping Sun
Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) emerges as a prospective therapeutic angle in regenerative medicine and a tool for drug screening. Although increasing numbers of iPSCs from different sources have been generated, there has been limited progress in yield of iPSC. Here, we show that four Yamanaka factors Oct4, Sox2, Klf4 and c-Myc can convert human embryonic renal cortical cells (hERCCs) to pluripotent stem cells with a roughly 40-fold higher reprogramming efficiency compared with that of adult human dermal fibroblasts...
2016: American Journal of Translational Research
https://www.readbyqxmd.com/read/27903888/sirna-carrying-an-e-vinylphosphonate-moiety-at-the-5-end-of-the-guide-strand-augments-gene-silencing-by-enhanced-binding-to-human-argonaute-2
#8
Elad Elkayam, Rubina Parmar, Christopher R Brown, Jennifer L Willoughby, Christopher S Theile, Muthiah Manoharan, Leemor Joshua-Tor
Efficient gene silencing by RNA interference (RNAi) in vivo requires the recognition and binding of the 5'- phosphate of the guide strand of an siRNA by the Argonaute protein. However, for exogenous siRNAs it is limited by the rapid removal of the 5'- phosphate of the guide strand by metabolic enzymes. Here, we have determined the crystal structure of human Argonaute-2 in complex with the metabolically stable 5'-(E)-vinylphosphonate (5'-E-VP) guide RNA at 2.5-Å resolution. The structure demonstrates how the 5' binding site in the Mid domain of human Argonaute-2 is able to adjust the key residues in the 5'-nucleotide binding pocket to compensate for the change introduced by the modified nucleotide...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27903806/oschx14-is-involved-in-the-k-homeostasis-in-rice-oryza-sativa-flowers
#9
Yi Chen, Jingkun Ma, Anthony J Miller, Bingbing Luo, Mei Wang, Zhen Zhu, Pieter B F Ouwerkerk
Previously we showed in the osjar1 mutants that the lodicule senescence which controls the closing of rice flowers was delayed. This resulted in florets staying open longer when compared with the wild type. The gene OsJAR1 is silenced in osjar1 mutants and is a key member of the jasmonic acid (JA) signaling pathway. We found that K concentrations in lodicules and flowers of osjar1-2 were significantly elevated compared with the wild type, indicating that K(+) homeostasis may play a role in regulating the closure of rice flowers...
July 2016: Plant & Cell Physiology
https://www.readbyqxmd.com/read/27903072/a-5-non-coding-exon-containing-engineered-intron-enhances-transgene-expression-from-recombinant-aav-vectors-in-vivo
#10
Jiamiao Lu, James A Williams, James Luke, Feijie Zhang, Kirk Chu, Mark A Kay
We previously developed a mini-intronic plasmid (MIP) expression system in which the essential bacterial elements for plasmid replication and selection are placed within an engineered intron contained within a universal 5'UTR non-coding exon. Like minicircle DNA plasmids (devoid of bacterial backbone sequences), MIP plasmids overcome transcriptional silencing of the transgene. However, in addition MIP plasmids increase transgene expression by 2 and often >10 times higher than minicircle vectors in vivo and in vitro...
November 30, 2016: Human Gene Therapy
https://www.readbyqxmd.com/read/27902457/tfdp3-confers-chemoresistance-in-minimal-residual-disease-within-childhood-t-cell-acute-lymphoblastic-leukemia
#11
Ming Chu, Kailin Yin, Yujun Dong, Pingzhang Wang, Yun Xue, Peng Zhou, Yuqi Wang, Yuedan Wang
Acquired drug resistance in childhood T-cell acute lymphoblastic leukemia (T-ALL) remains a significant clinical problem. In this study, a novel gene therapy target for childhood T-ALL to overcome chemoresistance was discovered: TFDP3 increased in the minimal residual disease (MRD) positive childhood T-ALL patients. Then, we established a preclinical model of resistance to induction therapy to examine the functional relevance of TFDP3 to chemoresistance in MRD derived from Jurkat/E6-1. Jurkat xenografts in NOD/SCID mice were exposed to a four drug combination (VXLD) of vincristine (VCR), dexamethasone (DEX), L-asparaginase (L-asp) and daunorubicin (DNR)...
November 26, 2016: Oncotarget
https://www.readbyqxmd.com/read/27901317/dendrimer-stabilized-gold-nanostars-as-a-multifunctional-theranostic-nanoplatform-for-ct-imaging-photothermal-therapy-and-gene-silencing-of-tumors
#12
Ping Wei, Jingwen Chen, Yong Hu, Xin Li, Han Wang, Mingwu Shen, Xiangyang Shi
Development of versatile nanomaterials combining diagnostic and therapeutic functionalities within one single nanoplatform is extremely important for tumor theranostics. In this work, the authors report the synthesis of a gold nanostar (Au NS)-based theranostic platform stabilized with cyclic arginine-glycine-aspartic (Arg-Gly-Asp, RGD) peptide-modified amine-terminated generation 3 poly(amidoamine) dendrimers. The formed RGD-modified dendrimer-stabilized Au NSs (RGD-Au DSNSs) are used as a gene delivery vector to complex small interfering RNA (siRNA) for computed tomography (CT) imaging, thermal imaging, photothermal therapy (PTT), and gene therapy of tumors...
November 30, 2016: Advanced Healthcare Materials
https://www.readbyqxmd.com/read/27899818/bnc2-is-a-putative-tumor-suppressor-gene-in-high-grade-serous-ovarian-carcinoma-and-impacts-cell-survival-after-oxidative-stress
#13
Laura Cesaratto, Eleonora Grisard, Michela Coan, Luigi Zandonà, Elena De Mattia, Elena Poletto, Erika Cecchin, Fabio Puglisi, Vincenzo Canzonieri, Maria Teresa Mucignat, Antonella Zucchetto, Gabriele Stocco, Alfonso Colombatti, Milena S Nicoloso, Riccardo Spizzo
Rs3814113 is the single-nucleotide polymorphism (SNP) showing the strongest association with high-grade serous ovarian carcinoma (HGSOC) incidence and is located in an intergenic region about 44 kb downstream of basonuclin 2 (BNC2) gene. Lifetime number of ovulations is associated with increased risk to develop HGSOC, probably because of cell damage of extrauterine Müllerian epithelium by ovulation-induced oxidative stress. However, the impact of low-penetrance HGSOC risk alleles (e.g. rs3814113) on the damage induced by oxidative stress remains unclear...
September 22, 2016: Cell Death & Disease
https://www.readbyqxmd.com/read/27899448/stk40-functions-as-a-novel-regulator-of-skeletal-muscle-differentiation
#14
Ke He, Jing Hu, Hongyao Yu, Lina Wang, Fan Tang, Junjie Gu, Laixiang Ge, Hongye Wang, Sheng Li, Ping Hu, Ying Jin
Skeletal muscle differentiation is a precisely coordinated process and the molecular mechanism regulating the process remains incompletely understood. Here, we report the identification of serine/threonine kinase 40 (Stk40) as a novel positive regulator of skeletal myoblast differentiation in culture and fetal skeletal muscle formation in vivo. We show that the expression level of Stk40 increases during skeletal muscle differentiation. Down-regulation and overexpression of Stk40 significantly decreases and increases myogenic differentiation of C2C12 myoblasts, respectively...
November 29, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27898347/human-amniotic-epithelial-cells-inhibit-cd4-t-cell-activation-in-acute-kidney-injury-patients-by-influencing-the-mir-101-c-rel-il-2-pathway
#15
Junfeng Liu, Rong Hua, Zhangbin Gong, Bin Shang, Yongyi Huang, Lihe Guo, Te Liu, Jun Xue
In the pathogenesis of acute kidney injury (AKI), the release of multiple interleukins can lead to increased kidney damage. Human amniotic epithelial cells (HuAECs) can inhibit immune cell activation in vivo and in vitro. We hypothesized that HuAECs could weaken patient-derived peripheral blood CD4+ T-cell activation and decreasing the ability of these cells to express and release IL-2. -Cell proliferation assay revealed that under the same culture conditions, activated AKI patient-derived CD4+ T cells had a significantly reduced proliferation rate when were co-cultured with HuAECs...
November 26, 2016: Molecular Immunology
https://www.readbyqxmd.com/read/27895746/rnai-mediated-gene-silencing-of-vascular-endothelial-growth-factor-c-suppresses-growth-and-induces-apoptosis-in-mouse-breast-cancer-in-vitro-and-in-vivo
#16
Yong-Chao Liu, Wen-Hui Ma, Yin-Lin Ge, Mei-Lan Xue, Zheng Zhang, Jin-Yu Zhang, Lin Hou, Run-Hong Mu
Vascular endothelial cell growth factor (VEGF)-C promotes tumorigenesis by allowing lymph node metastasis and lymphangiogenesis, among other actions. RNA interference (RNAi) is a novel technique for suppressing target gene expression and may increase the effectiveness of cancer treatments. The present study assessed the influence of VEGF-C RNAi on the apoptosis and proliferation of mouse breast cancer cells in vitro and in vivo. A total of three pairs of small interfering RNA (siRNA) targeting mouse VEGF-C were designed and synthesized prior to transfection into 4T1 cells via a liposomal approach...
November 2016: Oncology Letters
https://www.readbyqxmd.com/read/27893850/a-convenient-in-vivo-model-using-small-interfering-rna-silencing-to-rapidly-assess-skeletal-gene-function
#17
Wen Zhang, Can Liu, Bao Hai, Guohong Du, Hong Wang, Huijie Leng, Yingsheng Xu, Chunli Song
It is difficult to study bone in vitro because it contains various cell types that engage in cross-talk. Bone biologically links various organs, and it has thus become increasingly evident that skeletal physiology must be studied in an integrative manner in an intact animal. We developed a model using local intraosseous small interfering RNA (siRNA) injection to rapidly assess the effects of a target gene on the local skeletal environment. In this model, 160-g male Sprague-Dawley rats were treated for 1-2 weeks...
2016: PloS One
https://www.readbyqxmd.com/read/27891609/role-of-trpm7-in-cerebral-ischaemia-and-hypoxia
#18
Hong-Shuo Sun
Transient receptor potential melastatin 7 (TRPM7) channel, a calcium-permeable non-selective divalent cation channel, is broadly expressed in various cells and tissues, including the brain. TRPM7 is thought to be coupled to the metabolic state and regulate calcium homeostasis in the cell. TRPM7 takes part in a wide range of cell biology processes that affect cell growth and proliferation, as well as in embryonic development and skeleton formation. TRPM7 plays a significant role in ischaemic and hypoxic brain injury and neuronal cell death...
November 27, 2016: Journal of Physiology
https://www.readbyqxmd.com/read/27891591/nor1-suppresses-cancer-stem-like-cells-properties-of-tumor-cells-via-the-inhibition-of-the-akt-gsk-3%C3%AE-wnt-%C3%AE-catenin-aldh1a1-signal-circuit
#19
Wei Wang, Mei Yi, Shengnan Chen, Junjun Li, Haijing Zhang, Wei Xiong, Guiyuan Li, Xiaoling Li, Bo Xiang
Cancer stem cells (CSCs) play a key role in tumor radiotherapy and chemotherapy resistance, relapse, and metastasis, and are primarily maintained in a resting state in vivo. The failure of conventional therapies to target CSCs is the main cause of treatment failure. The discovery of CSCs in nasopharyngeal carcinoma (NPC) tumors is becoming more prevalent; however, the understanding of the mechanisms underlying the maintenance of tumor stemness is still limited. We previously cloned NOR1, a tumor suppressor gene downregulated in NPC cell lines and tissues...
November 27, 2016: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/27885273/fatty-acid-binding-protein-fabp4-mechanistically-links-obesity-with-aggressive-aml-by-enhancing-aberrant-dna-methylation-in-aml-cells
#20
F Yan, N Shen, J X Pang, Y W Zhang, E Y Rao, A M Bode, A Al-Kali, D E Zhang, M R Litzow, B Li, S J Liu
Obesity is becoming more prevalent worldwide and is a major risk factor for cancer development. Acute myeloid leukemia (AML), the most common acute leukemia in adults, remains a frequently fatal disease. Here we investigated the molecular mechanisms by which obesity favors AML growth and uncovered the fatty acid-binding protein 4 (FABP4) and DNA methyltransferase 1 (DNMT1) regulatory axis that mediates aggressive AML in obesity. We showed that leukemia burden was much higher in high-fat diet-induced obese mice, which had higher levels of FABP4 and interleukin (IL)-6 in the sera...
December 2, 2016: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
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