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Antisense oligodeoxynucleotides are typically targeted to bind mRNA sequences, leading to inhibition of gene expression by activation of RNase H to cleave the mRNA, obstruction of translation, alteration of splicing, or other mechanisms. The experimental determination of an effective antisense DNA to inhibit the expression of a particular gene product is expensive and time-consuming, and efforts have long been made to develop a procedure for the rational design of antisense DNA sequences based on properties such as the DNA:RNA hybrid stability, the region of the mRNA being targeted, and the secondary structures of the mRNA and DNA (reviewed by Chan et al. [1]). Programs using in vitro thermodynamic information for intrastrand and interstrand DNA and RNA interactions can be used to help discriminate weak from potent antisense DNA sequences [2, 3]. While extremely important for understanding stabilities of base pairs in vitro, the underlying thermodynamic information in such programs (e.g. the ...
TY - JOUR. T1 - Efficient delivery of an antisense oligodeoxyribonucleotide formulated in folate receptor-targeted liposomes. AU - Chiu, Shih Jiuan. AU - Marcucci, Guido. AU - Lee, Robert J.. PY - 2006/3. Y1 - 2006/3. N2 - Background: Folate receptors (FRs) are cellular surface markers for numerous solid tumors and myeloid leukemias. The aim of this study was to develop an antisense oligodeoxyribonucleotide (ODN) carrier targeting FR-overexpressing cancer cells using folate (FA) as the targeting moiety. G3139, a phosphorothioate antisense ODN against human bcl2 mRNA, was evaluated in this study. Materials and Methods: G3139-containing liposomes were prepared using an ethanol dilution method. For the targeted formulation, 0.5 mol% of folate-PEG-DSPE was incorporated as a targeting ligand into cationic liposomes composed of DC-Chol/egg PC/PEG-DSPE at 25:65:10 mol/mol. Particle size and surface charge were measured and cellular uptake was assessed by fluorescence microscopy and flow cytometry. The ...
Recent studies had found thousands of natural antisense transcripts originating from the same genomic loci of protein coding genes but from the opposite strand. It is unclear whether the majority of antisense transcripts are functional or merely transcriptional noise. Using the Affymetrix Exon array with a modified cDNA synthesis protocol that enables genome-wide detection of antisense transcription, we conducted large-scale expression analysis of antisense transcripts in nine corresponding tissues from human, mouse and rat. We detected thousands of antisense transcripts, some of which show tissue-specific expression that could be subjected to further study for their potential function in the corresponding tissues/organs. The expression patterns of many antisense transcripts are conserved across species, suggesting selective pressure on these transcripts. When compared to protein-coding genes, antisense transcripts show a lesser degree of expression conservation. We also found a positive correlation
Antisense oligonucleotides are an emerging therapeutic option to treat diseases with known genetic origin. In the age of personalised medicines, antisense oligonucleotides can sometimes be designed to target and bypass or overcome a patient's genetic mutation, in particular those lesions that compromise normal pre-mRNA processing. Antisense oligonucleotides can alter gene expression through a variety of mechanisms as determined by the chemistry and antisense oligomer design. Through targeting the pre-mRNA, antisense oligonucleotides can alter splicing and induce a specific spliceoform or disrupt the reading frame, target an RNA transcript for degradation through RNaseH activation, block ribosome initiation of protein translation or disrupt miRNA function. The recent accelerated approval of eteplirsen (renamed Exondys 51™) by the Food and Drug Administration, for the treatment of Duchenne muscular dystrophy, and nusinersen, for the treatment of spinal muscular atrophy, herald a new and exciting era in
TY - JOUR. T1 - Phase II study of ISIS 3521, an antisense oligodeoxynucleotide to protein kinase C alpha, in patients with previously treated low-grade non-Hodgkin's lymphoma. AU - Rao, S.. AU - Watkins, D.. AU - Cunningham, D.. AU - Dunlop, D.. AU - Johnson, P.. AU - Selby, P.. AU - Hancock, B.W.. AU - Fegan, C.. AU - Culligan, D.. AU - Schey, S.. AU - Morris, T. AU - Lissitchkov, T.. AU - Oliver, J.W.. AU - Holmlund, J.T.. PY - 2004/1. Y1 - 2004/1. N2 - Background: The purpose of this study was to assess the efficacy and safety of ISIS 3521, an antisense phosphorothioate oligonucleotide to protein kinase C in patients with relapsed low-grade non-Hodgkin's lymphoma (NHL). Patients and methods: Twenty-six patients received ISIS 3521 (2 mg/kg/day) as a continuous infusion over 21 days of each 28-day cycle. Results: The median age of the patients was 53 years (range 37-77). Histological subtypes were low-grade follicular lymphoma (n=22) and B-cell small lymphocytic lymphoma (n=4). Twenty-one (81%) ...
An important new collection of clinical and preclinical reports on genetic therapy, this book describes illustrative examples of diseases in which gene-based interventions are presently plausible, and presents case studies of current research usingMoreAn important new collection of clinical and preclinical reports on genetic therapy, this book describes illustrative examples of diseases in which gene-based interventions are presently plausible, and presents case studies of current research using both synthetic oligonucleotides and biological vectors.Combining the insights of over 50 contributors, Clinical Trials of Genetic Therapy with Antisense DNA and DNA Vectorsfurnishes a historical overview of genetic therapyhighlights official Food and Drug Administration positions on the preparation of oligonucleotides and vectorsoffers practical models of agent preparation, animal testing, pharmacokinetics, toxicology, and clinical trialsdiscusses both synthetic DNA and biological vector approaches to ...
article{4ac00774-2f85-4776-9b6d-b14fe0384556, abstract = {Antisense oligodeoxynucleotides to endothelin ETA receptor mRNA were used to characterize vascular smooth muscle receptors. The concentration-response curve showed a significant attenuation of endothelin-1-induced contraction in circular segments of the human superficial temporal artery. Endothelin ETB receptor antisense or mismatch oligodeoxynucleotides showed no alteration of the endothelin-1-induced contraction. Complementary experiments with the selective endothelin ETA receptor antagonist FR139317 demonstrated a shift of the concentration-response curve to the right in a competitive manner (pA2 = 6.93). The specific method of using the receptor antisense oligodeoxynucleotides approach revealed the presence of endothelin ETA receptors mediating contraction in the human superficial temporal artery.}, author = {Adner, Mikael and Erlinge, David and Salford, Leif and Yee, Frances and Wahlestedt, Claes and Edvinsson, Lars}, issn = ...
Despite the advances in our understanding of transcriptome, regulation and function of its noncoding components continue to be poorly understood. Here we searched for natural antisense transcript for sensorin (NAT-SRN), a neuropeptide expressed in the presynaptic sensory neurons of gill-withdrawal reflex of the marine snail Aplysia californica. Sensorin (SRN) has a key role in learning and long-term memory storage in Aplysia. We have identified NAT-SRN in the central nervous system (CNS) and have confirmed its expression by northern blotting and fluorescent RNA in situ hybridization. Quantitative analysis of NAT-SRN in micro dissected cell bodies and processes of sensory neurons suggest that NAT-SRN is present in the distal neuronal processes along with sense transcripts. Importantly, aging is associated with reduced levels of NAT-SRN in sensory neuron processes. Furthermore, we find that forskolin, an activator of CREB signaling, differentially alters the distribution of SRN and NAT-SRN. These studies
Erwinia amylovora is a Gram-negative bacterial plant pathogen in the family Enterobacteriaceae and is the causal agent of fire blight, a devastating disease of apple and pear. Fire blight is traditionally managed by the application of the antibiotic streptomycin during bloom, but this strategy has been challenged by the development and spread of streptomycin resistance. Thus, there is an urgent need for effective, specific, and sustainable control alternatives for fire blight. Antisense antimicrobials are oligomers of nucleic acid homologs with antisense sequence of essential genes in bacteria. The binding of these molecules to the mRNA of essential genes can result in translational repression and antimicrobial effect. Here, we explored the possibility of developing antisense antimicrobials against E. amylovora and using these compounds in fire blight control. We determined that a 10-nucleotide oligomer of peptide nucleic acid (PNA) targeting the start codon region of an essential gene acpP is able to
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Primer recognition proteins (PRP) are cofactors for DNA polymerase alpha and may have a role in lagging-strand DNA replication. PRP is composed of two subunits, which we have previously identified as the protein-tyrosine kinase substrate annexin II and phosphoglycerate kinase (PGK). In this study, we have examined the physiological involvement of these proteins in DNA synthesis and cell proliferation. When exponentially growing human HeLa cells are exposed to antisense phosphorothioate oligodeoxynucleotides to annexin II, ongoing DNA synthesis is reduced. The extent of reduction with antisense oligodeoxynucleotide to PGK was much less than with the antisense annexin II oligodeoxynucleotide. Reductions in the labeling and mitotic indices of HeLa cell cultures are seen after exposure to antisense oligodeoxynucleotides. Flow cytometric analyses indicate that progression from S phase to G2 phase of the cycle is retarded by exposure of cells to the antisense oligodeoxynucleotides. Corresponding sense ...
Walter Reed Army Institute of Research 1964-1966. John Mark Freeman was born in Brooklyn, New York, and educated at Deerfield Academy and Amherst College prior to studying medicine and completing a residency in pediatrics at Johns Hopkins. His decision to become a child neurologist was based on the particular influence of David Clark. Dr. Freeman's interactions with Arnold Gold, Sidney Carter, John Menkes, and Horace Hodes awakened the interest in research that has so strongly marked his career development. Dr. Freeman was awarded Columbia's Lucy Moses prize for research in neurology.. After two years of research at Walter Reed and then three years on the faculty at Stanford, he returned to Johns Hopkins as director of the child neurology service and of the birth defects treatment center. During the 21 years that he spent in. these, he rose to the rank of professor and in 1991 was named the Lederer Professor of Pediatric Epilepsy. His particular concentration in epilepsy was enriched by ...
Texas health-care worker tests positive for Ebola. An image from the U.S. Centres for Disease Control shows an Ebola Virus.. Phase one of the trials will focus on determining the proper dosage and any possible side effects of the vaccine.. Twenty vials of the vaccine have been sent to the Walter Reed Army Institute of Research in Maryland for testing on about 40 healthy volunteers, she said.. The announcement comes after a health worker in Texas became the first person to be infected with Ebola inside the United States. The nurse contracted it from a patient, a Liberian man who developed symptoms shortly after arriving in U.S.. American health officials say a breach of infection protocol led to the nurse catching the disease.. The Winnipeg Regional Health Authority has ordered additional supplies as part of their ongoing preparations for a possible Ebola diagnosis here.. When asked if the WRHA is concerned about the infection of Texas health worker, a spokesperson said the agency has sent ...
L-lysyl-L-threonyl-L phenylalanyl-N-3-carboxypropyl)-glycine amide, acetate salt. 5) Treatment of perinatal asphyxia - Allopurinol sodium. 6) Treatment of Duchenne muscular dystrophy - Exon 52 specific phosphorothioate oligonucleotide. 7) Treatment of Duchenne muscular dystrophy - Exon 55 specific phosphorothioate oligonucleotide. 8) Treatment of malaria - Artesunate. 9) Treatment of chronic lymphocytic leukaemia - 4-(4-{[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl}piperazin-1-yl)-N-({3-nitro-4-[(tetrahydro-2H-pyran-4-ylmethyl)amino]phenyl}sulfonyl)-2-(1H-pyrrolo[2,3-b]pyridin-5-yloxy)benzamide. 10) Treatment of very-long-chain-acyl-CoA dehydrogenase deficiency - Triheptanoin. 11) Treatment of long-chain L-3-hydroxyacyl-CoA-dehydrogenase deficiency - Triheptanoin. 12) Treatment of chronic lymphocytic leukaemia - Humanised single chain monoclonal antibody against CD37. 13) Treatment of non-infectious uveitis - Voclosporin. Copyright © 2012-2013, Orphan Druganaut Blog. All rights ...
PURPOSE: To examine the signal transduction pathways involved in the activation of orbital fibroblast effector functions relevant to the pathogenesis of Graves' ophthalmopathy (GO). To determine, using antisense technology, whether the c-myc protooncogene is involved in cell proliferation and glycosaminoglycan (GAG) synthesis in cultured orbital fibroblasts (OF). METHODS: The effects of a 16-mer c-myc antisense phosphorothioate oligodeoxynucleotide (S-ODN) on OF monolayers derived from orbital connective tissue of patients with severe GO (n = 6) and healthy individuals (n = 3) were investigated. Quiescent OF monolayers were treated with serum or cytokines and were exposed to increasing concentrations of a c-myc antisense S-ODN and several control S-ODN. Cell proliferation was quantitated by direct cell counting and by immunocytochemistry for the nuclear Ki-67 antigen. Glycosaminoglycan synthesis was examined by [3H] GAG analysis. The effects of the c-myc antisense S-ODN and control S-ODN on ...
Antibiotic resistance genes can be targeted by antisense agents, which can reduce their expression and thus restore cellular susceptibility to existing antibiotics. Antisense inhibitors can be gene and pathogen specific, or designed to inhibit a group of bacteria having conserved sequences within resistance genes. Here, we aimed to develop antisense peptide nucleic acids (PNAs) that could be used to effectively restore susceptibility to β-lactams in methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP). Antisense PNAs specific for conserved regions of the mobilisable gene mecA, and the growth essential gene, ftsZ, were designed. Clinical MRSA and MRSP strains of high oxacillin resistance were treated with PNAs and assayed for reduction in colony forming units on oxacillin plates, reduction in target gene mRNA levels, and cell size. Anti-mecA PNA at 7.5 and 2.5 μM reduced mecA mRNA in MRSA and MRSP (p | 0.05). At these PNA concentrations, 66
The amyloid-beta (A beta) peptide is a cytotoxic peptide implicated in the pathology of Alzheimer's disease (AD). Catalase and the endoplasmic reticulum A beta binding dehydrogenase (ERAB) are both inhibited by characterized fragments of the A beta peptide. In order to target such proteins it is ess …
Author: Stitt, M. et al.; Genre: Journal Article; Published in Print: 1991; Keywords: flux control (photosynthesis)|br/|nicotiana (transformed with antisense DNA)|br/|ribulose-1,5-bisphosphate carboxylase-oxygenase (control of photosynthesis)|br/|transgenic plant (antisense)|br/|c-3 plants|br/|leaves|br/|limitations|br/|metabolism|br/|phosphate|br/|nitrogen|br/|fixation; Title: Decreased Ribulose-1,5-Bisphosphate Carboxylase-Oxygenase in Transgenic Tobacco Transformed with Antisense Rbcs. 2. Flux-Control Coefficients for Photosynthesis in Varying Light, Co2, and Air Humidity
Baculovirus IAP (inhibitor-of-apoptosis) genes originated by capture of host genes. Unmodified short antisense DNA oligonucleotides (oligoDNAs) from baculovirus IAP genes can down-regulate specific gene expression profiles in both baculovirus-free and baculovirus-infected insects. In this study, gypsy moth (Lymantria dispar) larvae infected with multiple nucleopolyhedrovirus (LdMNPV), and LdMNPV-free larvae, were treated with oligoDNA antisense to the RING (really interesting new gene) domain of the LdMNPV IAP-3 gene. The results with respect to insect mortality, biomass accumulation, histological studies, RT-PCR, and analysis of DNA apoptotic fragmentation suggest that oligoRING induced increased apoptotic processes in both LdMNPV-free and LdMNPV-infected insect cells, but were more pronounced in the latter. These data open up possibilities for promising new routes of insect pest control using antisense phosphodiester DNA oligonucleotides.
BACKGROUND: X chromosome inactivation is the mechanism used in mammals to achieve dosage compensation of X-linked genes in XX females relative to XY males. Chromosome silencing is triggered in cis by expression of the non-coding RNA Xist. As such, correct regulation of the Xist gene promoter is required to establish appropriate X chromosome activity both in males and females. Studies to date have demonstrated co-transcription of an antisense RNA Tsix and low-level sense transcription prior to onset of X inactivation. The balance of sense and antisense RNA is important in determining the probability that a given Xist allele will be expressed, termed the X inactivation choice, when X inactivation commences. RESULTS: Here we investigate further the mechanism of Xist promoter regulation. We demonstrate that both sense and antisense transcription modulate Xist promoter DNA methylation in undifferentiated embryonic stem (ES) cells, suggesting a possible mechanistic basis for influencing X chromosome choice.
Antisense Elements (Antisense Genetic Elements) are Nucleic acids which hybridize to complementary sequences in other target nucleic acids causing the function of the latter nucleic acids to be affected
Genomic imprinting is a process that leads to the silencing of one allele of a gene in a parent of origin specific manner. Genes that are involved in this process are often regulated in clusters, one of which is the Peg3 (Paternally expressed gene 3) imprinted domain. We investigated this region for both CpG islands and long antisense transcripts, two common features of imprinted gene clusters. First, we performed a systematic survey of DNA methylation status of the CpG islands in this region of the mouse, cow, and human genomes. We identified two previously unreported differentially methylated regions (DMR): one in the promoter region of mouse Zim3 and another in the promoter region of human USP29. The PEG3-CpG island is the only DMR that is conserved among these three species. PEG3 has been implicated in several types of cancer, so we examined the methylation status of several CpG islands in this region using human tumor derived DNA. The CpG islands near PEG3 and USP29 both showed hypermethylation in
James Franklin Crow (January 18, 1916 - January 4, 2012) was Professor Emeritus of Genetics at the University of Wisconsin-Madison and a prominent population geneticist whose career spanned from the modern synthesis to the genomic era. Some of his most significant peer-reviewed contributions were coauthored with Motoo Kimura, including those leading to the neutral theory of molecular evolution. He also wrote an influential introductory textbook on genetics and a more advanced one with Kimura. His graduate and undergraduate students and postdocs includes Alexey Kondrashov, James Bull, Joe Felsenstein, Russell Lande, Dan Hartl, and Wen-Hsiung Li. He was a president of both the Genetics Society of America and the American Society of Human Genetics. He was a member of the National Academy of Sciences, The American Philosophical Society, the World Academy of Art and Science, the National Academy of Medicine, the American Academy of Arts and Sciences, and Foreign Member of the Royal Society ...
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Antisense oligonucleotides directed at the bcl-xl gene product engender apoptosis in mesothelioma cell lines. The therapeutic potential of inhibiting expression of this protein in mesothelioma should be evaluated.
Orotic Acid and/or Orotidine may be elevated secondary to hyperammonemia. Increased excretion of Orotic Acid is seen in Ornithine Transcarbamylase (OTC) Deficiency as well as Hereditary Orotic Aciduria ...
The antiviral potential of transcripts targeted to the non-coding regions (NCRs) of foot-and-mouth disease virus (FMDV) RNA have been studied during transient and constitutive expression in susceptible BHK-21 cells. Transient expression of antisense transcripts corresponding to the 5′ and 3′NCRs, alone or in combination, confers specific inhibition of homologous (serotype C) virus infection in BHK-21 cells. Constitutive expression of antisense 5′NCR transcripts (5′AS) exerted higher levels of inhibition to homologous and heterologous (serotypes O, A, Asia, SAT 1, SAT 2 and SAT 3) FMDV infection, as estimated by a 10-fold reduction in virus titre in the supernatants from infected clones and by a plaque reduction assay. These inhibitions were also observed, albeit to a lesser extent, in clones stably expressing antisense 3′NCR transcripts. The antiviral response was specific for FMDV, as the picornavirus encephalomyocarditis virus was not inhibited in any of the transformed cell lines. In all
The potential for therapeutic application of splice-switching oligonucleotides (SSOs) to modulate pre-mRNA splicing is increasingly evident in a number of diseases. However, the primary drawback of this approach is poor cell and in vivo oligonucleotide uptake efficacy. Biological activities can be significantly enhanced through the use of synthetically conjugated cationic cell penetrating peptides (CPPs). Studies to date have focused on the delivery of a single SSO conjugated to a CPP, but here we describe the conjugation of two phosphorodiamidate morpholino oligonucleotide (PMO) SSOs to a single CPP for simultaneous delivery and pre-mRNA targeting of two separate genes, exon 23 of the Dmd gene and exon 5 of the Acvr2b gene, in a mouse model of Duchenne muscular dystrophy. Conjugations of PMOs to a single CPP were carried out through an amide bond in one case and through a triazole linkage ('click chemistry') in the other. The most active bi-specific CPP-PMOs demonstrated comparable exon skipping levels
TY - JOUR. T1 - Down-regulation of type I protein kinase A by transfection of human breast cancer cells with an epidermal growth factor receptor antisense expression vector. AU - Ciardiello, Fortunato. AU - Dixit, Mniralini. AU - Di Isernia, Giuditta. AU - Damiano, Vincenzo. AU - Bianco, Roberto. AU - Bianco, A. Raffaele. AU - Arteaga, Carlos L.. AU - Tortora, Giampaolo. PY - 1998/2/5. Y1 - 1998/2/5. N2 - MDA-468 human breast cancer cells overexpress the EGFR and exhibit a functional TGFα-EGFR autocrine pathway. Loss of EGFR expression following stable transfection with an antisense EGFR cDNA containing plasmid down-regulates type T cAMP-dependent protein kinase (PKAI) expression with acquisition of cell growth resistance to the PKAI inhibitor 8-Cl-cAMP. These results suggest that PKAI expression and function are controlled by a TGFα-EGFR autocrine pathway in human breast cancer cells overexpressing the EGFR.. AB - MDA-468 human breast cancer cells overexpress the EGFR and exhibit a functional ...
Holmes SE, O'Hearn E, Callahan C, Hwang HS, Rosenblatt A, Ingersoll-Ashworth RG, Fleisher A, Stevanin G, Brice A, Potter NT, Ross CA, Margolis RL. A CTG trinucleotide repeat expansion in Junctophilin 3 is associated with Huntington's Disease-Like 2 (HDL2). Nature Genetics, 29 (2001): 377-378, 2001.. Chung DW, Rudnicki DD, Yu L, Margolis RL. A natural antisense transcript at the Huntington's disease repeat locus regulates HTT expression. Human Molecular Genetics, 20 (2011):3467-77.. Chiang C-H, Su Y, Wen Z, Yoritomo N, Ross CA, Margolis RL (co-corresponding author), Song H, Ming G-l. Integration-free induced pluripotent stem cells derived from schizophrenia patients with a DISC1 mutation. Molecular Psychiatry 16 (2011):358-60.. Seixas AI, Holmes SE, Takeshima H, Pavlovich A, Sachs N, Pruitt JL, Silveira I, Ross CA, Margolis RL, Rudnicki DD. Loss of junctophilin-3 contributes to Huntington's Disease-like 2 pathogenesis, Annals of Neurology, 71(2012):245-257.. Yu L, Arbez N, Nucifora LG, Sells G, ...
TY - JOUR. T1 - Exon skipping and dystrophin restoration in patients with Duchenne muscular dystrophy after systemic phosphorodiamidate morpholino oligomer treatment: An open-label, phase 2, dose-escalation study. AU - Cirak, Sebahattin. AU - Arechavala-Gomeza, Virginia. AU - Guglieri, Michela. AU - Feng, Lucy. AU - Torelli, Silvia. AU - Anthony, Karen. AU - Abbs, Stephen. AU - Garralda, Maria Elena. AU - Bourke, John. AU - Wells, Dominic J.. AU - Dickson, George. AU - Wood, Matthew Ja. AU - Wilton, Steve D.. AU - Straub, Volker. AU - Kole, Ryszard. AU - Shrewsbury, Stephen B.. AU - Sewry, Caroline. AU - Morgan, Jennifer E.. AU - Bushby, Kate. AU - Muntoni, Francesco. PY - 2011/8/13. Y1 - 2011/8/13. N2 - We report clinical safety and biochemical efficacy from a dose-ranging study of intravenously administered AVI-4658 phosphorodiamidate morpholino oligomer (PMO) in patients with Duchenne muscular dystrophy. We undertook an open-label, phase 2, dose-escalation study (0·5, 1·0, 2·0, 4·0, ...
AstraZeneca has launched a phase IIb clinical trial of the investigational antisense agent, ION449 (AZD8233), in patients with dyslipidemia.. The drug is developed by AstraZeneca in partnership with California-based biotech company Ionis Pharmaceuticals. Ionis has received a milestone payment of $20m from AstraZeneca for the initiation of the trial.. ION449 is a ligand conjugated antisense (LICA) medicine designed to lower blood cholesterol levels by targeting proprotein convertase subtilisin/kexin type 9 (PCSK9). PCSK9 is an enzyme that controls the number of low-density lipoprotein cholesterol (LDL-C) receptors on the surface of cells.. The randomised, double-blind, placebo-controlled trial will enrol around 108 adults aged between 18 and 75 with LDL-C levels between 70 and 190 mg/dL and are receiving moderate or high-intensity statin therapy.. The effect of different doses of ION449 on LDL-C versus placebo at week 12 in patients taking baseline statin therapy will be the trial's primary ...
TY - JOUR. T1 - Phosphorothioate-modified oligodeoxynucleotides inhibit human cytomegalovirus replication by blocking virus entry. AU - Luganini, Anna. AU - Caposio, Patrizia. AU - Landolfo, Santo. AU - Gribaudo, Giorgio. PY - 2008/3. Y1 - 2008/3. N2 - Studies in animal models have provided evidence that Toll-like receptor 9 (TLR9) agonists, such as synthetic oligodeoxynucleotides (ODNs) that contain immunostimulatory deoxycytidyl-deoxyguanosine (CpG) motifs (CpG ODNs), protect against a wide range of viral pathogens. This antiviral activity has been suggested to be indirect and secondary to CpG-induced cytokines and inflammatory responses triggered through TLR9 activation. However, few studies have addressed the potential of CpG ODNs as direct antiviral agents. Here, we report on the ability of some CpG ODNs to directly suppress, almost completely, human cytomegalovirus (HCMV) replication in both primary fibroblasts and endothelial cells. Murine CMV replication was inhibited as well, whereas no ...
Abstract: : Purpose: To determine the effect of phosphorothioate antisense oligonucletides directed against c-met/HGF receptor on RPE cell differentiation and proliferation. Methods: We transfected cultured human RPE with c-met-specific phosphorothioate antisense oligonucleotides that were designed as 18-mer sequences complementary to c-met mRNA. Target mRNA was quantified by RNase protection assay. Cellular differentiation and proliferation were determined by nitro blue tetrazolium (NBT) dye reduction and immunocytochemistry of CD44 RPE cell surface antigen expression. Results: Transfection of c-met antisense oligonucleotides (2uM) reduced the expression of c-met mRNA by 50% to 65%. It also inhibited RPE cell proliferation and blocked CD44 expression. Conclusion: Antisense oligonucleotides effectively inhibit c-met expression, RPE cell differentiation and proliferation. ...
The general transcription factor TATA-binding protein (TBP) is a key initiation factor involved in transcription by all three eukaryotic RNA polymerases. In addition, the related metazoan-specific TBP-like factor (TLF/TRF2) is essential for transcription of a distinct subset of genes. Here we characterize the vertebrate-specific TBP-like factor TBP2, using in vitro assays, in vivo antisense knockdown, and mRNA rescue experiments, as well as chromatin immunoprecipitation. We show that TBP2 is recruited to promoters in Xenopus oocytes in the absence of detectable TBP recruitment. Furthermore, TBP2 is essential for gastrulation and for the transcription of a subset of genes during Xenopus embryogenesis. In embryos, TBP2 protein is much less abundant than TBP, and moderate overexpression of TBP2 partially rescues an antisense knockdown of TBP levels and restores transcription of many TBP-dependent genes. TBP2 may be a TBP replacement factor in oocytes, whereas in embryos both TBP and TBP2 are ...
Antonio MANCARELLA (from February 2014). The genome of HIV-1 harbors the three common retroviral genes (gag, pol, and env), in addition to two regulatory genes (tat and rev) and four accessory genes (vif, vpr, vpu, and nef). All of these genes are expressed through a single transcript initiating from the 5' long terminal repeat (LTR), in which the promoter is located. The various viral proteins are synthesized from unspliced, monospliced, or multispliced forms of this major transcript and then assume their functional roles in infected cells.. Recently, several data have suggested that another gene, encoding a product named HIV antisense protein (ASP), may be expressed through an antisense transcript. Antisense transcription encoding proteins involved in the modulation of transactivation potential of multiple cellular transcription activators has been shown in HTLV. Antisense transcription has also been suggested for HIV-1, based on the identification of conserved ORFs in the antisense strand of ...
http://www.ncbi.nlm.nih.gov/pubmed/19074264. Bestman and Cline used their in vivo single-cell electroporation protocol to introduce lissamine-tagged Morpholino antisense oligos into individual optic tectal neurons in stage 46 to 48 albino Xenopus laevis tadpoles. They knocked down expression of cytoplasmic polyadenylation element binding protein 1 (CPEB1), a component of some brain ribonucleoprotein (RNP) granules. CPEB1 binds to RNA that have cytoplasmic polyadenylation elements (CPE) in their 3'-untranslated regions; CPEB1 is estimated by bioinformatics to bind to about 7% of brain mRNAs. Binding of CPEB1 to an mRNA's CPE represses its translation and regulates microtubule-dependant trafficking. When CPEB1 is phosphorylated, the mRNA can be polyadenylated and translated.. The authors collected time-lapse images of neurons over a period of three days after treatment with either CPEB1-targeted Morpholinos or control Morpholinos. In either treatment the dendritic arbors grew, but the arbors of ...
We have shown that in human cells, an antiserum generated against a recombinant UBPY fragment recognizes a protein doublet of Mr 130 000, in good agreement with the predicted UBPY mol. wt of 127 500 Da. The UBPY bands were enhanced upon transfection of a sense UBPY cDNA and, most importantly, were lowered in abundance upon transfection of an antisense cDNA construct. UBPY was able to remove ubiquitin from ubiquitin adducts. Its expression was undetectable upon serum starvation of normal human fibroblasts, and the protein reappeared upon re‐stimulation, in mid G1 coincident with the accumulation of cyclin D1. UBPY levels were reduced in non‐immortalized cells as they reached confluence and arrested in G0, while they remained high and even increased in transformed cells.. To date, no other studies have demonstrated the effects of down‐regulating a UBP in mammalian cells. We have been able to inhibit UBPY accumulation using an antisense cDNA vector and could demonstrate that G0‐arrested ...
BACKGROUND: Targeted splice modulation of pre-mRNA transcripts by antisense oligonucleotides (AOs) can correct the function of aberrant disease-related genes. Duchenne muscular dystrophy (DMD) arises as a result of mutations that interrupt the open-reading frame in the DMD gene encoding dystrophin such that dystrophin protein is absent, leading to fatal muscle degeneration. AOs have been shown to correct this dystrophin defect via exon skipping to yield functional dystrophin protein in animal models of DMD and also in DMD patients via intramuscular administration. To advance this therapeutic method requires increased exon skipping efficiency via an optimized AO sequence, backbone chemistry and additional modifications, and the improvement of methods for evaluating AO efficacy. METHODS: In the present study, we establish the conditions for rapid in vitro AO screening in H(2)K muscle cells, in which we evaluate the exon skipping properties of a number of known and novel AO chemistries [2'-O-methyl,
The egg yolk of vertebrates contains carotenoids, which account for its characteristic yellow color in some species. Such plant-derived compounds, e.g. beta-carotene, serve as the natural precursors (provitamins) of vitamin A, which is indispensable for chordate development. As egg yolk also contains stored vitamin A, carotenoids have so far been solely discussed as pigments for the coloration of the offspring. Based on our recent molecular identification of the enzyme catalyzing provitamin A conversion to vitamin A, we address a possible role of provitamin A during zebrafish (Danio rerio) development. We cloned the zebrafish gene encoding the vitamin A-forming enzyme, a beta,beta-carotene-15,15'-oxygenase. Analysis of its mRNA expression revealed that it is under complex spatial and temporal control during development. Targeted gene knockdown using the morpholino antisense oligonucleotide technique indicated a vital role of the provitamin A-converting enzyme. Morpholino-injected embryos ...
Kennedy's disease is a neurodegenerative disease caused by a gene mutation which encodes the androgen receptor. The signs and symptoms are clearly visible among patients. However with a prevalence of only 1 in 30.000 patients worldwide the disease is classified as rare. Research to date has shown no cure for ... read more this trinucleotide (CAG) repeat disorder which attacks the motor neurons and muscles. Small molecule drugs and biologics have failed to be beneficial and antisense technology should make a difference. Basically this approach offers new opportunities for neurodegenerative disorders especially Kennedy disease. In co-operation with Utrecht University, University of Glasgow and Prosensa we want to look at RNA modulation for Kennedy's disease. The purpose of this research project is to synthesize and investigate several single stranded antisense oligonucleotides, with the antisense mRNA sequence 3' UUG-(CUGX)-CAG 5', where X stands for 10, 20, 30, 40, 50 or 60 CAG repeats (normally: ...
Antisense transcription is widespread in genomes. Despite large differences in gene size and architecture, we find that yeast and human genes share a unique, antisense transcription-associated chromatin signature. We asked whether this signature is related to a biological function for antisense transcription. Using quantitative RNA-FISH, we observed changes in sense transcript distributions in nuclei and cytoplasm as antisense transcript levels were altered. To determine the mechanistic differences underlying these distributions, we developed a mathematical framework describing transcription from initiation to transcript degradation. At GAL1, high levels of antisense transcription alter sense transcription dynamics, reducing rates of transcript production and processing, while increasing transcript stability. This relationship with transcript stability is also observed as a genome-wide association. Establishing the antisense transcription-associated chromatin signature through disruption of the Set3C
Hoxa 11 is a murine Abdominal-B-type homeobox gene. The structure of this gene is presented, including genomic and cDNA sequence. The cDNA includes the complete open reading frame and based on primer extension results is near full length. Surprisingly, the antisense strand of Hoxa 11 was found to be transcribed. Moreover, these antisense transcripts were processed and polyadenylated. The developmental expression patterns for both sense and antisense transcripts were examined using serial section and whole-mount in situ hybridizations. Hoxa 11 transcription patterns were defined in the limbs, kidney and stromal cells surrounding the Mullerian and Wolffian ducts. Of particular interest, in the developing limbs, the sense and antisense transcripts showed complementary expression patterns, with antisense RNAs increasing in abundance in regions where sense RNAs were diminishing in abundance. Furthermore, targeted mutation of Hoxa 11 is shown to result in both male and female sterility. The female ...
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The abundance of long non-coding RNAs (lncRNAs) and their wide range of functional roles in human cells are fast becoming realized. Importantly, lncRNAs have been identified as epigenetic modulators and consequently play a pivotal role in the regulation of gene expression. A human immunodeficiency virus (HIV) encoded antisense RNA transcript has recently been reported and researchers at the Scripps Research Institute sought to characterize this RNA and determine its potential role in viral transcription regulation. The intrinsic properties of this HIV-expressed lncRNA were characterized and the data presented here suggest that it functions as an epigenetic brake to modulate viral transcription. Suppression of this long antisense transcript with small single stranded antisense RNAs resulted in the activation of viral gene expression. This lncRNA was found to localize to the 5'LTR and to usurp components of endogenous cellular pathways that are involved in lncRNA directed epigenetic gene ...
N-myc proto-oncogene protein also known as N-Myc or basic helix-loop-helix protein 37 (bHLHe37), is a protein that in humans is encoded by the MYCN gene. The MYCN gene is a member of the MYC family of transcription factors and encodes a protein with a basic helix-loop-helix (bHLH) domain. This protein is located in the cell nucleus and must dimerize with another bHLH protein in order to bind DNA. N-Myc is highly expressed in the fetal brain and is critical for normal brain development. The MYCN gene has an antisense RNA, N-cym or MYCNOS, transcribed from the opposite strand which can be translated to form a protein product. N-Myc and MYCNOS are co-regulated both in normal development and in tumor cells, so it is possible that the two proteins are functionally related. It has been shown that NCYM antisense RNA encodes for a protein that has originated de novo and is specific to human and chimpanzee. This NCYM protein inhibits GSK3b and thus prevents MYCN degradation. Transgenic mice that harbor ...
The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of a Sirtuin (SIRT), in particular, by targeting natural antisense polynucleotides of a Sirtuin (SIRT). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of Sirtuins (SIRT)s.
Isis Publishes Data Demonstrating Antisense Targeting of ApoC-III Significantly Reduces ApoC-III and Triglycerides Research Shows Antisense Inhibition of ApoC-III and Triglycerides in Multiple...
In this report, we show how a convenient on-resin copper-click functionalization of azido-functionalized peptide nucleic acids (PNAs) allows various PNA-based detection strategies. Firstly, a thiazole orange (TO) clicked PNA probe facilitates a binary readout when combined with F/Q labeled DNA, giving increa