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In vivo gene silencing

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https://www.readbyqxmd.com/read/30244460/rna-interference-ex-vivo
#1
Shuang Liu
RNA interference (RNAi) is a widely used technique to regulate the expression of genes and proteins with a high degree of specificity that is not easily accessed by traditional pharmacological approaches. For preclinical research on rheumatoid arthritis (RA), silencing of target genes in primary immune cells can be easily achieved by application of small interfering RNA (siRNA) and synthetic short hairpin RNA (shRNA). Cellular and systemic administration of siRNA or shRNA has been a significant advance in preclinical research on RA...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/30244246/role-of-mir-148a-in-mitigating-hepatic-ischemia-reperfusion-injury-by-repressing-the-tlr4-signaling-pathway-via-targeting-camkii%C3%AE-in-vivo-and-in-vitro
#2
Daofeng Zheng, Zhongtang Li, Xufu Wei, Rui Liu, Ai Shen, Diao He, Chengyong Tang, Zhongjun Wu
BACKGROUND/AIMS: Hepatic ischemia-reperfusion (I/R) injury, which is mainly induced by inflammation and unstable intracellular ions, is a major negative consequence of surgery that compromises hepatic function. However, the exact mechanisms of liver I/R injury have not been determined. Positive crosstalk with the Ca2+/CaMKII pathway is required for complete activation of the TLR4 pathway and inflammation. We previously found that miR-148a, which decreased in abundance with increasing reperfusion time, targeted and repressed the expression of CaMKIIα...
September 21, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/30240228/fluorinated-oligoethylenimines-nano-assemblies-for-efficient-sirna-mediated-gene-silencing-in-serum-containing-media-by-effective-endosomal-escape
#3
Tingbin Zhang, Yuanyu Huang, Xiaowei Ma, Ningqiang Gong, Xiaoli Liu, Lu Liu, Xiaoxia Ye, Bo Hu, Chunhui Li, Jianhua Tian, Andrea Magrini, Jinchao Zhang, Weisheng Guo, Jinfeng Xing, Massimo Bottini, Xing-Jie Liang
Efficient small interfering RNA (siRNA) delivery in presence of serum is of crucial importance for effective gene therapy. Fluorinated vectors are considered to be attractive candidates for siRNA-mediated gene therapy because of their delivery efficacy in serum-containing media. However, the mechanisms driving the superior gene transfection behavior of fluorinated vectors are still not well understood and comprehensive investigations are warranted. Herein, we fabricated a library of perfluorooctanoyl fluoride (PFF)-fluorinated oligoethylenimines (fxOEIs, x is the PFF:OEI feeding ratio), which can readily form nano-assemblies (fxOEI NAs) capable of efficient siRNA delivery in cells cultured in medium both devoid of and supplemented with fetal bovine serum (FBS)...
September 21, 2018: Nano Letters
https://www.readbyqxmd.com/read/30237545/cpeb1-mediates-hepatocellular-carcinoma-cancer-stemness-and-chemoresistance
#4
Min Xu, Shiji Fang, Jingjing Song, Minjiang Chen, Qianqian Zhang, Qiaoyou Weng, Xiaoxi Fan, Weiqian Chen, Xulu Wu, Fazong Wu, Jianfei Tu, Zhongwei Zhao, Jiansong Ji
Cancer stem cells (CSCs) are a subpopulation of cells within tumors that are believed to possess pluripotent properties and thought to be responsible for tumor initiation, progression, relapse and metastasis. Cytoplasmic polyadenylation element-binding protein 1 (CPEB1), a sequence-specific RNA-binding protein that regulates mRNA polyadenylation and translation, has been linked to cancer progression and metastasis. However, the involvement of CPEB1 in hepatocellular carcinoma (HCC) remains unclear. In this study, we have demonstrated that CPEB1 directly regulates sirtuin 1 (SIRT1) mRNA to mediate cancer stemness in HCC...
September 20, 2018: Cell Death & Disease
https://www.readbyqxmd.com/read/30237402/female-mice-lacking-ftx-lncrna-exhibit-impaired-x-chromosome-inactivation-and-a-microphthalmia-like-phenotype
#5
Yusuke Hosoi, Miki Soma, Hirosuke Shiura, Takashi Sado, Hidetoshi Hasuwa, Kuniya Abe, Takashi Kohda, Fumitoshi Ishino, Shin Kobayashi
X-chromosome inactivation (XCI) is an essential epigenetic process in female mammalian development. Although cell-based studies suggest the potential importance of the Ftx long non-protein-coding RNA (lncRNA) in XCI, its physiological roles in vivo remain unclear. Here we show that targeted deletion of X-linked mouse Ftx lncRNA causes eye abnormalities resembling human microphthalmia in a subset of females but rarely in males. This inheritance pattern cannot be explained by X-linked dominant or recessive inheritance, where males typically show a more severe phenotype than females...
September 20, 2018: Nature Communications
https://www.readbyqxmd.com/read/30236833/hsa_circ_0061140-knockdown-reverses-foxm1-mediated-cell-growth-and-metastasis-in-ovarian-cancer-through-mir-370-sponge-activity
#6
Qizhen Chen, Jiarong Zhang, Yinyan He, Yanqiu Wang
Circular RNAs (circRNAs) are a class of noncoding RNAs that regulate gene expression at the posttranscriptional level. The specific functions of circRNAs in ovarian cancer are yet to be established. Previous sequencing analyses have revealed an abnormal expression of hsa_circ_0061140 in ovarian cancer. The main aim of the present study is to establish the specific role of hsa_circ_0061140 in ovarian cancer. circRNA expression in ovarian cancer cells was detected via real-time qPCR. The effects on specific cellular characteristics (proliferation, migration, and the EMT) and subcellular localization of hsa_circ_0061140 were assessed via RNA fluorescence in situ hybridization, knockdown, and luciferase reporter assays in the SKOV3 and A2780 cell lines...
August 22, 2018: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/30236487/the-regulatory-role-of-annexin-3-in-a-nude-mouse-bearing-a-subcutaneous-xenograft-of-mda-mb-231-human-breast-carcinoma
#7
Jie Li, Tao Zhou, Liang Liu, Ying Chao Ju, Yue Tong Chen, Zi Rui Tan, Jing Wang
The following study investigated the effects of Annexin A3 (ANXA3) on breast cancer biological behavior in vivo, using nude mouse model bearing a subcutaneous tumor. A total of 18 female nude mice were randomly divided into three groups (n = 6): negative control group which was inoculated with MDA-MB-231 cells, blank control group which was inoculated with MDA-MB-231-NC cells, and the transfection group which was inoculated with MDA-MB-231-Sh cells. The experiment lasted for 4 weeks, during which mice conditions, diet and defecation were monitored on a daily basis...
September 13, 2018: Pathology, Research and Practice
https://www.readbyqxmd.com/read/30233146/anti-her2-functionalized-graphene-oxide-as-survivin-sirna-delivery-carrier-inhibits-breast-carcinoma-growth-in-vitro-and-in-vivo
#8
Xiaoli Wang, Qi Sun, Chunying Cui, Jing Li, Yifan Wang
Background: The success of gene therapy is mostly dependent on the development of gene carrier. Graphene oxide (GO) possesses excellent aqueous solubility and biocompatibility, which is important for its biochemical and medical applications. Our previous work proved that GO can deliver siRNA into cells efficiently and downregulate the expression of desired protein. Methods: In this study, a novel delivery carrier, GO-R8/anti-HER2 (GRH), was developed by conjugating octaarginine (R8) and anti-HER2 antibody with GO as a tumor active-targeting vector for survivin-siRNA delivery...
2018: Drug Design, Development and Therapy
https://www.readbyqxmd.com/read/30232346/in-silico-and-in-vivo-analyses-of-the-mutated-human-tissue-plasminogen-activator-mtpa-and-the-antithetical-effects-of-p19-silencing-suppressor-on-its-expression-in-two-nicotiana-species
#9
Mahshid Amiri, Mokhtar Jalali-Javaran, Raheem Haddad, Parastoo Ehsani
Human tissue-type plasminogen activator is one of the most important therapeutic proteins involved in the breakdown of blood clots following the stroke. A mutation was found at position 1541 bp (G514E) and the mutated form was cloned into the binary vector pTRAc-ERH. In silico analysis showed that this mutation might have no significant effect on the active site of the tissue plasminogen activator enzyme. Accordingly, zymography assay confirmed the serine protease activity of the mutated form and its derivatives...
September 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/30229544/effective-inhibition-of-different-japanese-encephalitis-virus-genotypes-by-rna-interference-targeting-two-conserved-viral-gene-sequences-in-vitro-and-in-vivo
#10
Lei Yuan, Xiaojuan Feng, Xuelian Gao, Yu Luo, Chaoyue Liu, Peng Liu, Guolin Yang, Hong Ren, Rong Huang, Yalan Feng, Jian Yang
Japanese encephalitis is a zoonotic, mosquito-borne, infectious disease caused by Japanese encephalitis virus (JEV), which is prevalent in China. At present, there are no specific drugs or therapies for JEV infection, which can only be treated symptomatically. Lentivirus-mediated RNA interference (RNAi) is a highly efficient method to silence target genes. In this study, two lentiviral shRNA, LV-C and LV-NS5, targeting the conserved viral gene sequences were used to inhibit different JEV genotypes strains in BHK21 cells and mice...
September 18, 2018: Virus Genes
https://www.readbyqxmd.com/read/30227958/loss-of-deubiquitinase-usp1-blocks-pancreatic-%C3%AE-cell-apoptosis-by-inhibiting-dna-damage-response
#11
Kanaka Durga Devi Gorrepati, Blaz Lupse, Karthika Annamalai, Ting Yuan, Kathrin Maedler, Amin Ardestani
Impaired pancreatic β-cell survival contributes to the reduced β-cell mass in diabetes, but underlying regulatory mechanisms and key players in this process remain incompletely understood. Here, we identified the deubiquitinase ubiquitin-specific protease 1 (USP1) as an important player in the regulation of β-cell apoptosis under diabetic conditions. Genetic silencing and pharmacological suppression of USP1 blocked β-cell death in several experimental models of diabetes in vitro and ex vivo without compromising insulin content and secretion and without impairing β-cell maturation/identity genes in human islets...
March 23, 2018: iScience
https://www.readbyqxmd.com/read/30223276/repression-of-pdk1-and-lncrna-hotair-mediated-ezh2-gene-expression-contributes-to-the-enhancement-of-atractylenolide-1-and-erlotinib-in-the-inhibition-of-human-lung-cancer-cells
#12
Qian Xiao, Fang Zheng, Qing Tang, Jing-Jing Wu, Jianhui Xie, Hai-Ding Huang, Xiao-Bo Yang, Swei Sunny Hann
BACKGROUND/AIMS: We previously showed that the major bioactive compound of Atractylodes macrocephula Koidz atractylenolide 1 (ATL-1) inhibited human lung cancer cell growth by suppressing the gene expression of 3-Phosphoinositide dependent protein kinase-1 (PDK1 or PDPK1). However, the potentially associated molecules and downstream effectors of PDK1 underlying this inhibition, particularly the mechanism for enhancing the anti-tumor effects of epidermal growth factor receptor-tyrosine-kinase inhibitors (EGFR-TKIs), remain unknown...
September 17, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/30221032/a-ribonucleoprotein-octamer-for-targeted-sirna-delivery
#13
Wanyi Tai, Junwei Li, Eva Corey, Xiaohu Gao
Hurdles in cell-specific delivery of small interfering RNA (siRNA) in vivo hinder the clinical translation of RNA interference (RNAi). A fundamental problem concerns conflicting requirements for the design of the delivery vehicles: cationic materials facilitate cargo condensation and endosomolysis, yet hinder in vivo targeting and colloidal stability. Here, we describe a self-assembled, compact (~30 nm) and biocompatible ribonucleoprotein-octamer nanoparticle that achieves endosomal destabilization and targeted delivery...
2018: Nature Biomedical Engineering
https://www.readbyqxmd.com/read/30218368/experimental-analysis-of-imprinted-mouse-x-chromosome-inactivation
#14
Marissa Cloutier, Clair Harris, Srimonta Gayen, Emily Maclary, Sundeep Kalantry
X-chromosome inactivation is a dosage compensation mechanism that equalizes X-linked gene expression between male and female mammals through the transcriptional silencing of most genes on one of the two X-chromosomes in females. With a few key exceptions, once the X-chromosome is inactivated replicated copies of that X-chromosome are maintained as inactive in all descendant cells. X-inactivation is therefore a paradigm of epigenetic inheritance. Imprinted X-inactivation is a specialized form of X-inactivation that results in the silencing of the paternally derived X-chromosome...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/30217818/phosphatase-activity-of-small-c-terminal-domain-phosphatase-1-scp1-controls-the-stability-of-the-key-neuronal-regulator-re1-silencing-transcription-factor-rest
#15
Nathaniel Tate Burkholder, Joshua E Mayfield, Xiaohua Yu, Seema Irani, Daniel K Arce, Faqin Jiang, Wendy Matthews, Yuanchao Xue, Yan Jessie Zhang
The RE1-silencing transcription factor (REST) is the major scaffold protein for assembly of neuronal gene silencing complexes that suppress gene transcription through regulating the surrounding chromatin structure. REST represses neuronal gene expression in stem cells and non-neuronal cells, but it is minimally expressed in neuronal cells to ensure proper neuronal development. Dysregulation of REST function has been implicated in several cancers and neurological diseases. Modulating REST gene silencing is challenging since cellular and developmental differences can affect its activity...
September 14, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/30217697/overexpression-of-gp73-promotes-cell-invasion-migration-and-metastasis-by-inducing-epithelial-mesenchymal-transition-in-pancreatic-cancer
#16
Yin-Xue Song, Zhi-Chao Xu, Hui-Ling Li, Pei-Lei Yang, Jun-Kai Du, Jing Xu
Pancreatic cancer is one of the most difficult clinical cases to diagnose with a very low 5-year survival rate of 5%, regardless of the advances made in both the medical and surgical treatment of the disease. One of the contributing factors for the high mortality rate seen of pancreatic cancer patients is the lack of effective chemotherapies, which is believed to be due to drug-resistance. Based on recent evidence, epithelial-mesenchymal transition (ETM) of pancreatic cancer cells has been found to be associated with the development of drug resistance and an increase in cell invasion...
August 23, 2018: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
https://www.readbyqxmd.com/read/30215317/generation-of-footprint-free-canine-induced-pluripotent-stem-cells-using-auto-erasable-sendai-virus-vector
#17
Masaya Tsukamoto, Toshiya Nishimura, Kyohei Yodoe, Ryoji Kanegi, Yasunori Tsujimoto, Md Emtiaj Alam, Mizuki Kuramochi, Mitsuru Kuwamura, Manami Ohtaka, Ken Nishimura, Mahito Nakanishi, Toshio Inaba, Kikuya Sugiura, Shingo Hatoya
Canine induced pluripotent stem cells (ciPSCs) can be used in regenerative medicine. However, there are no reports on the generation of genome integration free and completely exogeneous gene silenced (footprint-free) ciPSCs that are tolerant to enzymatic single cell passage. In this study, we reprogrammed canine embryonic fibroblasts (CEFs) using the auto-erasable replication-defective and persistent Sendai virus vector, SeVdp(KOSM)302L and generated two ciPSC lines. The ciPSCs were positive for pluripotent markers including alkaline phosphatase activity as well as OCT3/4, SOX2 and NANOG transcripts and NANOG, Stage Specific Embryonic Antigen-1 and partial TRA-1-60 protein expression, even after SeVdp(KOSM)302L removal...
September 14, 2018: Stem Cells and Development
https://www.readbyqxmd.com/read/30214300/silencing-the-expression-of-copine-iii-enhances-the-sensitivity-of-hepatocellular-carcinoma-cells-to-the-molecular-targeted-agent-sorafenib
#18
Zhuo Chen, Zhengkui Jiang, Wenzhou Zhang, Baoxia He
Background: The application of the oral targeted therapeutic agent sorafenib provides new hope for patients suffering from advanced stages of hepatocellular carcinoma (HCC), but the prognosis of such patients remains poor due to the rapid development of the multidrug resistance process in cancer pathogenesis. The present work evaluated whether copine-III, a novel cancer regulator encoded by the CPNE3 gene, would be a potential indicator of sorafenib resistance in HCC treatment. Materials and methods: The endogenous expression of copine-III in clinical specimens was examined by quantitative polymerase chain reaction...
2018: Cancer Management and Research
https://www.readbyqxmd.com/read/30213224/the-notch-family-transcription-factor-rbpj%C3%AE%C2%BA-modulates-glucose-transporter-and-ovarian-steroid-hormone-receptor-expression-during-decidualization
#19
Michael R Strug, Ren-Wei Su, Tae Hoon Kim, Jae-Wook Jeong, Asgerally Fazleabas
During decidualization, endometrial stromal cells differentiate into a secretory phenotype to modulate the uterine microenvironment and promote embryo implantation. This highly metabolic process relies on ovarian steroid receptors and glucose transporters. Canonical Notch signaling is mediated by the transcription factor Recombination Signal Binding Protein for Immunoglobulin Kappa J Region (RBPJ). Loss of RBPJ in the mouse uterus ( Pgrcre/+ Rbpjflox/flox ; Rbpj c-KO) results in subfertility in part due to an abnormal uterine-embryonic axis during implantation and, as described herein, decidualization failure...
September 13, 2018: Reproductive Sciences
https://www.readbyqxmd.com/read/30210918/long-noncoding-rna-tob1-as1-an-epigenetically-silenced-gene-functioned-as-a-novel-tumor-suppressor-by-sponging-mir-27b-in-cervical-cancer
#20
Jihang Yao, Zhenghong Li, Ziwei Yang, Hui Xue, Hua Chang, Xue Zhang, Tianren Li, Kejun Guo
Cervical cancer is one of the most common cancers in females, accounting for a majority of cancer-related deaths in worldwide. Long non-coding RNAs (lncRNAs) have been identified as critical regulators in many tumor-related biological processes. Thus, investigation into the function and mechanism of lncRNAs in the development of cervical cancer is very necessary. In this study, we found that the expression of TOB1-AS1 was significantly decreased in cervical cancer tissues compared with the adjacent normal tissues...
2018: American Journal of Cancer Research
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