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 ...

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. ...

Background: Clusterin is a cytoprotective chaperone protein that promotes cell survival and confers broad-spectrum treatment resistance. OGX-011 is a 2′-methoxyethyl modified phosphorothioate antisense oligonucleotide that is complementary to clusterin mRNA and has been reported to inhibit clusterin expression and enhance drug efficacy in xenograft models. The primary objective of this clinical study was to determine a biologically effective dose of OGX-011 that would inhibit clusterin expression in human cancer. Methods: Subjects (n = 25) with localized prostate cancer with high-risk features who were candidates for prostatectomy were treated with OGX-011 by 2-hour intravenous infusion on days 1, 3, and 5 and then weekly from days 8-29 combined with androgen blockade starting on day 1; prostatectomy was performed on days 30-36. Six different doses were tested, from 40 to 640 mg. OGX-011 plasma and prostate tissue concentrations were measured by an enzyme-linked immunosorbent assay method, and ...

0019]An essential requirement in the antisense approach is that an oligonucleotide or its analogue recognize and bind tightly to its complementary target RNA. The ability of the resulting antisense oligomer/RNA hybrid to serve as a substrate of RNaseH is likely to have therapeutic value by enhancing the antisense effect relative to oligomers that are unable to activate this enzyme. Apart from PS-DNA (phosphorothioates), PS-DNA (phosphorodithioates), boranophosphonate-linked DNA, and MBO oligos containing an internal PS-DNA segment, there are no other examples of fully modified oligonucleotides that elicit RNaseH activity. For this reason, and because of the problems encountered with PS-oligonucleotides (e.g., non-antisense effects and potential risk of toxicity), we have designed alternative oligonucleotide analogues that selectively block gene expression through the activation of RNaseH activity. As a starting point, we felt that such analogues should (a) retain the natural β-D-furanose ...

Charcot-Marie-Tooth disease type 1A (CMT1A) is caused by duplication of peripheral myelin protein 22 (PMP22) and is the most common hereditary peripheral neuropathy. CMT1A is characterized by demyelination and axonal loss, which underlie slowed motor nerve conduction velocity (MNCV) and reduced compound muscle action potentials (CMAP) in patients. There is currently no known treatment for this disease. Here, we show that antisense oligonucleotides (ASOs) effectively suppress PMP22 mRNA in affected nerves in 2 murine CMT1A models. Notably, initiation of ASO treatment after disease onset restored myelination, MNCV, and CMAP almost to levels seen in WT animals. In addition to disease-associated gene expression networks that were restored with ASO treatment, we also identified potential disease biomarkers through transcriptomic profiling. Furthermore, we demonstrated that reduction of PMP22 mRNA in skin biopsies from ASO-treated rats is a suitable biomarker for evaluating target engagement in ...

The IAP survivin deserves attention as a target for cancer therapy due to its differential expression in tumors versus normal tissues. It is expressed during embryonal development, lacks expression in terminally differentiated adult tissues, and becomes reexpressed in transformed cell lines and a variety of human tumors, with highest levels being found in breast and lung cancer (1) . The expression of survivin in tumors is correlated with drug resistance and/or shorter survival of patients with non-small cell lung cancer (5) , colorectal cancer (7) , and neuroblastoma (9) . Despite recognition that survivin represents an attractive target for cancer therapy (2, 3, 4, 5, 6, 7, 8, 9, 10) , mainly survivin antisense cDNA fragments (11 , 12) and very recently also antisense oligonucleotides (13) have been used to elucidate the role of survivin during cell division and apoptosis. In the present study, we describe an antisense oligonucleotide approach to down-regulate survivin expression in the lung ...

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 ...

Antisense compounds, compositions and methods are provided for modulating the expression of C-reactive protein. The compositions comprise antisense compounds, particularly antisense oligonucleotides, targeted to nucleic acids encoding C-reactive protein. Methods of using these compounds for modulation of C-reactive protein expression and for treatment of diseases associated with expression of C-reactive protein are provided.

Here, we show that ApoER2 splicing is deregulated in Alzheimers disease in humans and correction of this defect with antisense oligonucleotides is therapeutic in mice. ApoER2 functions in signaling pathways that are critical for brain development and neuronal maintenance, which, when disrupted, lead to learning and memory deficits similar to those seen in Alzheimers disease (Beffert et al, 2005, 2006; Wasser et al, 2014). ApoER2 expression is regulated, in part, by alternative splicing of exon 19, which codes for a protein domain that is essential for signaling. Here, we demonstrate that the relative abundance of the ApoER2 isoform that includes exon 19 is lower in the brain of persons with AD compared to persons without cognitive impairment, suggesting that a decrease in active ApoER2 isoforms may be a hallmark of AD. We find a similar decrease in exon 19 inclusion in the brains of mice modeled to have AD. We identified a splicing‐related protein that inhibits exon 19 splicing and designed ...

Progress in our understanding of the molecular pathogenesis of human malignancies has provided therapeutic targets amenable to oligonucleotide (ON)-based strategies. Antisense ON-mediated splicing regulation in particular offers promising prospects since the majority of human genes undergo alternative splicing and since splicing defects have been found in many diseases. However, their implementation has been hampered so far by the poor bioavailability of nucleic acids-based drugs. Cell-penetrating peptides (CPPs) now appear as promising non-viral delivery vector for non-permeant biomolecules. We describe here new CPPs allowing the delivery of splice redirecting steric-block ON using either chemical conjugation or non-covalent complexation. We also describe a convenient and robust splice redirecting assay which allows the quantitative assessment of ON nuclear delivery.

Compositions and methods are provided for the treatment and diagnosis of influenza virus infections. In accordance with preferred embodiments, oligonucleotides and oligonucleotide analogs are provided which are specifically hybridizable with viral RNAs. The oligonucleotide comprises nucleotide units sufficient in identity and number to effect said specific hybridization. In other preferred embodiments, the oligonucleotides are specifically hybridizable with a transcription initiation site, a translation initiation site, 5-untranslated sequences, 3-untranslated sequences, and intron/exon junction of influenza virus mRNAs. In additional preferred embodiments, the oligonucleotides are specifically hybridizable with RNA sequences involved in splicing of the viral RNA, or in viral packaging. Methods of treating animals suffering from influenza virus infection are disclosed.

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

The chances of finding a cure for Angelman Syndrome, a rare genetic disorder that cause developmental delays in children, are looking better thanks to the development of antisense therapies. The disease advocacy group FAST, which is funding this research, is also driving education and awareness among parents.

Regulation of biological processes through the use of genetic elements is a central part of biological research and also holds great promise for future therapeutic applications. Oligonucleotides comprise a class of versatile biomolecules capable of modulating gene regulation. Gene therapy, the concept of introducing genetic elements in order to treat disease, presents a promising therapeutic strategy based on such macromolecular agents. Applications involving charged macromolecules such as nucleic acids require the development of the active pharmaceutical ingredient as well as efficient means of intracellular delivery. Cell-penetrating peptides are a promising class of drug delivery vehicles, capable of translocation across the cell membrane together with molecules otherwise unable to permeate cells, which has gained significant attention. In order to increase the effectiveness of cell-penetrating peptide-mediated delivery, further understanding of the mechanisms of uptake is needed in addition ...

Gene Tools, LLC is a limited liability company located in Philomath, Oregon, United States that manufactures Morpholino antisense oligos and delivery reagents. Gene Tools was founded in 1997 and began regularly shipping custom-sequence Morpholino oligos in 2000. Current products include Morpholino oligos, Vivo-Morpholinos (for improved delivery into cells) and Photo-Morpholinos (cleavable by 365nm light). The manager and general partner, Jim Summerton, is a pioneer in antisense research, conceived of and was co-inventor of the Morpholino antisense oligo structural type and founded the first antisense therapeutics company, Sarepta Therapeutics Inc. (formerly AntiVirals Inc., renamed AVI BioPharma Inc., renamed Sarepta Therapeutics Inc.). List of companies based in Oregon Summerton, JE (2005). Endo-porter: a novel reagent for safe, effective delivery of substances into cells. Ann N Y Acad Sci. 1058: 62-75. doi:10.1196/annals.1359.012. PMID 16394126. Morcos, PA (2001). Achieving efficient ...

TY - JOUR. T1 - Potent and nontoxic antisense oligonudeotides containing locked nucleic acids. AU - Wahlestedt, Claes. AU - Salmi, Peter. AU - Good, Liam. AU - Kela, Johanna. AU - Johnsson, Thomas. AU - Hökfelt, Tomas. AU - Broberger, Christian. AU - Porreca, Frank. AU - Lai, Josephine. AU - Ren, Kunkun. AU - Ossipov, Michael. AU - Koshkin, Alexei. AU - Jakobsen, Nana. AU - Skouv, Jan. AU - Oerum, Henrik. AU - Jacobsen, Mogens Havsteen. AU - Wengel, Jesper. N1 - Copyright: Copyright 2018 Elsevier B.V., All rights reserved.. PY - 2000/5/9. Y1 - 2000/5/9. N2 - Insufficient efficacy and/or specificity of antisense oligonucleotides limit their in vivo usefulness. We demonstrate here that a high-affinity DNA analog, locked nucleic acid (LNA), confers several desired properties to antisense agents. Unlike DNA, LNA/DNA copolymers were not degraded readily in blood serum and cell extracts. However, like DNA, the LNA/DNA copolymers were capable of activating RNase H, an important antisense mechanism of ...

Demonstramos a síntese de nanopartículas de silício poroso fusogenic para entrega efectiva do oligonucleotide in vitro e in vivo....

In article ,57e912$jfb at kadri.ut.ee,, mspeek at tamm.eenet.ee says... , ,Elizabeth Rosenberg (erosenbe at gpu2.srv.ualberta.ca) wrote: ,: Hi! , ,: Our group is about to embark on the glorious journey of downregulation of ,: a gene by use of antisense constructs. We are very new to this, and would ,: appreciate any and all feedback anyone might have. ,: Now we would like to insert an antisense fragment of the same gene into ,: the pREP10 vector and tranfect this construct into normal CHO cells that ,: are the parent of our mutant strain. ,I would not recommend antisense experiments such as yours. This is because ,it is remarkably inefficient (for review see NAR 21:4383-91 (1993)) ,I spent 1.5 yrs to study RNA-RNA hybridization/annealing between ,a pair of naturally occurring sense-antisense mRNAs, but found ,very little of them in duplex (Mol. Endo.9:1655-65 (1995)) ,Mart (mspeek at ebc.ee in Estonia) I would also think that you may need several, if not many very good controls. I have seen ...

Small interfering (si) RNAs and antisense oligonucleotides (ASOs; here for simplicity reasons, both referred to as oligonucleotides) are small synthetic RNA or DNA molecules with a sequence complementary to a (pre)mRNA. Although the basic mechanisms of action between siRNAs and ASO are distinct, a sequence-specific interaction of the both oligonucleotides with the target (pre)mRNA alters the targets fate, which includes highly effective sequence-specific blockade of translation and consequently depletion of the corresponding protein. For a number of years, these oligonucleotides have been used as a tool in biological research to study gene function in vitro. More recently, safe and specific delivery of these oligonucleotides to the liver of mammals has been achieved and optimized. This not only allowed their use for in vivo gene studies in physiology and disease, but also opened the opportunity for the development of a new generation of RNA-specific drugs for therapeutic purposes. In 2013, the ...

Antisense oligonucleotides (also known as ASOs and AONs) are small pieces of DNA designed to correct specific genetic mutations. Dr. Lentz and her team developed an ASO to correct the c.216,A mutation in the gene harmonin, which causes Usher syndrome type 1C (USH1C) in people with Acadian heritage. She demonstrated that her ASO treatment, delivered four times a year, improved harmonin expression in mice with the mutation. However, it is not clear if the level of improved expression would be sufficient to save or restore vision in humans with that mutation. She plans to evaluate other ASOs with the hope of increasing the production of the normal harmonin protein ...

Endogenous and xenobiotic-enhanced oxidative stress may initiate embryonic death and birth defects via reactive oxygen species (ROS) signaling pathways involving nuclear transcription factor-κB (NF-κB). Using embryo culture and a transgenic mouse engineered with a NF-κB-dependent β-galactosidase reporter gene, we employed NF-κB antisense oligonucleotide therapy to determine whether NF-κB signaling contributes to the embryopathic effects of the ROS-initiating teratogen phenytoin. Phenytoin selectively increased NF-κB activity in target tissues and caused embryopathies, both of which were blocked by NF-κB antisense oligonucleotides but not by sense and nonsense oligonucleotide controls. NF-κB signaling may therefore contribute to the mechanism of ROS-mediated embryopathies.. ...

Mutations that cause activation of the KRAS oncogene are common in human cancer, including treatment-resistant tumor types such as lung and pancreatic cancer. KRAS has also proven to be notoriously difficult to target with small molecules. To overcome this issue, Ross et al. have turned to genetic technology, demonstrating an antisense oligonucleotide-based therapy for inhibiting KRAS. The antisense oligonucleotide used in this study was chemically modified, allowing systemic delivery through subcutaneous injection and avoiding the need for a specialized delivery vehicle. The authors tested the efficacy of this therapy in multiple mouse models of non-small cell lung cancer and evaluated its safety in primates, demonstrating its potential suitability for translation to humans. ...

Structural abnormalities of the c-abl proto-oncogene are found in hematopoietic cells of more than 90 percent of individuals with chronic myelogenous leukemia. Therefore c-abl may be important in normal as well as malignant hematopoiesis. Normal human hematopoietic progenitor cells were exposed to three different c-abl sense or antisense oligodeoxynucleotides, and the effects on myeloid and erythroid colony formation were examined. The c-abl antisense oligodeoxynucleotides inhibited myeloid, but not erythroid, colony formation. The c-abl sense oligodeoxynucleotides and bcr sense and antisense oligodeoxynucleotides were not inhibitory in this assay. These data show that c-abl is critical in normal myelopoiesis and may explain the relatively selective expansion of leukocytes in patients with chronic myelogenous leukemia. ...

Another substance class on the rise in cardiovascular medicine are so-called anti-sense oligonucleotides, single strands of DNA or RNA binding complementary to a chosen mRNA sequence, thereby preventing protein translation. Besides a fascinating novel anti-coagulatory approach by inhibiting coagulation factor XI production, the biggest focus of this novel therapeutic approach lies on lipidology. Within this review we will highlight the current evidence on antisense therapy against apolipoprotein B, apolipoprotein A as well as apolipoprotein CIII, that are in very different stages of development, however, with some exciting early data.. Kurzfassung: Biologika begr nden eine neue Medikamentenklasse, werden biotechnologisch hergestellt und greifen gezielt in molekularbiologische Mechanismen ein. Lange Zeit eine Dom ne der H matologie, Onkologie sowie der Rheumatologie, treten monoklonale Antik rper als Therapeutika nun auch langsam in der Kardiologie ihren Siegeszug an. Neben dem schon seit mehr ...

AbstractIn the present study, a twenty-mer antisense oligonucleotide specific for N-methyl-D-aspartate receptor one (ANR1) was applied to striatal neurons in primary cell culture. The ANR1 was found to be specific and nontoxic. Significant reductions in expression of NR1 mRNA and proteins were resulted after a single dose of ANR1 transcripts. Interestingly, there were reductions in total NR1 proteins but two phosphorylated forms of NR1 proteins at serine 896 and 897 residues were not reduced. There was also no change in the pattern of distribution of NR1 immunoreactivity in the striatal neurons. In addition, significant reductions of NMDA-mediated peak inward current were found after application of a higher concentration of ANR1 (20-100 mu M) by patch clamp recordings. The present results indicate that ANR1 is a useful agent in reducing NMIDA receptor functions. The present data thus provide detailed cellular and molecular mechanisms to explain our previous findings of amelioration of motor ...

module Grid GK5 - Winter School}. The venue took place on 13 to 17 of January 2019 in Karolinska Instituet in Stockholm. It was organised by DARTER with the great help of {tip image=images/Participants/300px/RogerS.jpg}Prof.{/tip} Roger Strömberg, the host of the DARTER Winter School. The lectures focused on nucleic acid chemistry (basic reactions of nucleic acids and their components, synthesis of oligonucleotides, common modifications and oligonucleotide conjugates), oligonucleotide (ON) therapy overview, and basic structures of nucleic acids. It also covered some specific topics in ON therapy: splice-switching therapy and Duchenne muscular dystrophy, Triple repeat diseases, siRNA therapy, mRNA therapy, exosomes, antisense ON therapy, delivery, tissue targeting and industrial aspects.. ...

Gene Tools makes Morpholino antisense oligos. Morpholino oligos bind to complementary RNA and get in the way of processes; they can knock down gene expression, modify RNA splicing or inhibit miRNA activity and maturation. Morpholinos are the premier knockdown tools used in developmental biology labs, the best RNA-blocking reagents for cells in culture and, as Vivo-Morpholinos, the most specific delivery-enhanced oligos available for other animal models. We are the sole commercial manufacturer selling research quantities of Morpholinos world-wide. Morpholino oligos are short chains of about 25 Morpholino subunits. Each subunit is comprised of a nucleic acid base, a methylenemorpholine ring and a non-ionic phosphorodiamidate intersubunit linkage. Morpholinos do not degrade their RNA targets, but instead act via an RNAse H-independent steric blocking mechanism. They are completely stable in cells, uncut by nucleases. Requiring greater complementarity with their target RNAs to affect gene ...

Chemical modifications of oligonucleotides provide an important tool to understand how the natural substrate works as well as how to improve their biochemical and biological properties as potential therapeutics and diagnostics. Our carba-LNA (2,4-carba-bridged Locked Nucleic Acid) modified oligo-DNA or -RNA have been found to be useful to modulate oligo-RNA and -DNA activity. This thesis is based on four papers: Paper I (J. Org. Chem. 2010, 75, 7112-7128) deals with the synthesis of 2,4-propylene-bridged (Carba-ENA) thymidine and its analogues. These carba-ENA nucleosides have been subsequently incorporated into 15mer antisense oligodeoxynucleotides (AON), and their affinity toward complementary mRNA and DNA, as well as their nuclease resistance and RNase H recruitment capability have been investigated in comparison with those of the native and ENA counterparts. Paper II (J. Org. Chem. 2012, 77, 6855-6872) illustrates the synthesis of dimethylbicyclo[2.2.1]heptane and a diastereomeric ...

Natural antisense RNAs are endogenous molecules that are complementary to RNA transcripts of already established function. They were discovered first in prokaryotes in which they are now recognised as an important component of molecular mechanisms involved in the regulation of gene expression. Recently, through the cumulative efforts of molecular biologists and bioinformaticians, natural antisense RNAs have been demonstrated in significant numbers in eukaryotic systems also. Probably the most exciting outcome of these studies is that natural antisense RNAs are particularly prevalent in the nervous system. Here we discuss the major known types of natural antisense RNAs in eukaryotic systems and focus on their potential roles in the regulation of gene expression in the brain.. ...

The goal of this clinical research study is to find the highest safe dose of BP1001 , a liposomal Growth Factor Receptor Bound Protein-2 antisense oligodeoxynucleotide (L-Grb2 AS), that can be given as treatment for patients with Ph+ CML, AML, ALL, or MDS. BP1001 is an antisense drug, which means it may help stop cancer cells by blocking the action of a protein that signals cancer cells to divide and increase in number. The protein that BP1001 blocks is called Grb-2. The response of the leukemia to this treatment will also be studied. In addition, the time needed for the body to process this drug will be evaluated.

Activating mutations in KRAS underlie the pathogenesis of up to 20% of human tumors, and KRAS is one of the most frequently mutated genes in cancer. Developing therapeutics to block KRAS activity has proven difficult, and no direct inhibitor of KRAS function has entered clinical trials. We describe the preclinical evaluation of AZD4785, a high-affinity constrained ethyl-containing therapeutic antisense oligonucleotide (ASO) targeting KRAS mRNA. AZD4785 potently and selectively depleted cellular KRAS mRNA and protein, resulting in inhibition of downstream effector pathways and antiproliferative effects selectively in KRAS mutant cells. AZD4785-mediated depletion of KRAS was not associated with feedback activation of the mitogen-activated protein kinase (MAPK) pathway, which is seen with RAS-MAPK pathway inhibitors. Systemic delivery of AZD4785 to mice bearing KRAS mutant non-small cell lung cancer cell line xenografts or patient-derived xenografts resulted in inhibition of KRAS expression in ...

Chronic lymphocytic leukemia (CLL) and small B-cell lymphocytic lymphoma (SLL) are thought to be different manifestations of the same disease. Treatment options for CLL/SLL range from a watch and wait approach to bone marrow transplant. Currently there is no consensus on the best treatment regimen and new approaches to treatment are needed.. EL625 is a 20-mer antisense molecule which binds to a coding region of exon 10 in p53 RNA transcripts. It can bind to both mutant and wild type p53. p53 is involved in regulating apoptosis and DNA repair in cells. When genetic damage occurs p53 is upregulated. As the expression of p53 increases in normal cells they are more likely to undergo apoptosis rather than cell cycle arrest and DNA repair. However in malignant cells, for a given level of DNA damage they are more likely to undergo cell cycle arrest and repair rather than apoptosis. Because EL625 is theorized to increase response to chemotherapy, we propose adding EL625 to a combination of fludarabine, ...

The pharmacokinetics of a 2′-O-(2-methoxyethyl)-ribose modified phosphorothioate oligonucleotide, ISIS 104838 (human tumor necrosis factor-α antisense), have been characterized in mouse, rat, dog, monkey, and human. Plasma pharmacokinetics after i.v. administration exhibited relatively rapid distribution from plasma to tissues with a distribution half-life estimated from approximately 15 to 45 min in all species. Absorption after s.c. injection was high (80-100%), and absorption after intrajejunal administration in proprietary formulations was as high as 10% bioavailability compared with i.v. administration. Urinary excretion of the parent drug was low, with less than 1% of the administered dose excreted in urine after i.v. infusion in monkeys at clinically relevant doses (≤5 mg/kg). ISIS 104838 is highly bound to plasma proteins, likely preventing renal filtration. However, shortened oligonucleotide metabolites of ISIS 104838 lose their affinity to bind plasma proteins. Thus, excretion of ...

HOUSTON, June 28, 2016-- Bio-Path Holdings, Inc.,, a biotechnology company leveraging its proprietary DNAbilize™ liposomal delivery and antisense technology to develop a portfolio of targeted nucleic acid drugs, today announced that it has entered into a sponsored research agreement with Thomas Jefferson University to investigate DNAbilize™ antisense DNA...

The cationic lipid Genzyme lipid (GL) 67 is the current gold-standard for in vivo lung gene transfer. Here, we assessed, if GL67 mediated uptake of siRNAs and asODNs into airway epithelium in vivo. Anti-lacZ and ENaC (epithelial sodium channel) siRNA and asODN were complexed to GL67 and administered to the mouse airway epithelium in vivo Transfection efficiency and efficacy were assessed using real-time RT-PCR as well as through protein expression and functional studies. In parallel in vitro experiments were carried out to select the most efficient oligonucleotides. In vitro, GL67 efficiently complexed asODNs and siRNAs, and both were stable in exhaled breath condensate. Importantly, during in vitro selection of functional siRNA and asODN we noted that asODNs accumulated rapidly in the nuclei of transfected cells, whereas siRNAs remained in the cytoplasm, a pattern consistent with their presumed site of action. Following in vivo lung transfection siRNAs were only visible in alveolar macrophages,

AIMS: Cyclin-dependent kinase inhibitors (CDKIs) play a critical role in negatively regulating the proliferation of cardiomyocytes, although their role in cardiac differentiation remains largely undetermined. We have shown that the most prominent CDKI in Xenopus, p27(Xic1)(Xic1), plays a role in neuronal and myotome differentiation beyond its ability to arrest the cell cycle. Thus, we investigated whether it plays a similar role in cardiomyocyte differentiation. METHODS AND RESULTS: Xenopus laevis embryos were sectioned, and whole-mount antibody staining and immunofluorescence studies were carried out to determine the total number and percentage of differentiated cardiomyocytes in mitosis. Capped RNA and/or translation-blocking Xic1 morpholino antisense oligonucleotides (Xic1Mo) were microinjected into embryos, and their role on cardiac differentiation was assessed by in situ hybridization and/or PCR. We show that cell-cycling post-gastrulation is not essential for cardiac differentiation in ...

Introduction: Locked nucleic acid antisense oligonucleotides (LNA-ONs) potently down regulate mRNA expression in transfected cells (IC50 ~ 0.5 - 5.0 nM). While this class of molecules clearly has in vivo activity when given alone, systemic delivery of LNA-ONs may be further improved if more favorable pharmacokinetic profile, cell penetration, and specific organ targeting were possible. Improvement in the specific tumor targeting in vivo will practically increase the possibility of using oligonucleotide based molecules to treat a broad spectrum of human diseases. We report here the novel releasable PEG-nanoparticles that enhance accumulation of LNA-ON in solid tumors and improve the cellular delivery in tumors in mice. Experimental procedures: a series of PEG-lipids with releasable linkers were prepared and formulated with oligonucleotides to form nanoparticles. The nanoparticles were incubated under different pH and temperatures to study their stability. Anti-HER3 LNA-ON and anti-bcl2 siRNA were ...

Descriptive info: Homepage.. Welcome to BioSpring.. BioSpring , The Oligo Company.. Expertise in oligonucleotide synthesis.. Flexibility by open source oligonucleotide production system.. Synthesis of oligonucleotides.. Standard analyses of oligonucleotides.. Additional analyses of oligonucleotides.. Superstructures: Biological significance and structure.. siRNA, RNAi.. Interfering RNA.. Hybridisation of siRNA molecules.. Quality and delivery time.. Phosphonoacetates.. Antisense molecules.. RNase H activity.. Transport into the cells.. Advantages of phosphonoacetates.. Scales and Modifications.. Unmodified oligonucleotides.. Phosphorothioates.. RNA.. Modified RNA.. Methylphosphonates.. Phosphonoacetate oligonucleotides (PACE).. RNAi, siRNA miRNA.. 5 modifications.. 3 modifications.. Internal modifications.. Dual labeled probes.. Additional services.. Delivery.. Qualification and validation.. BioSpring GMP Manufacturing movie.. Oligonucleotides for diagnostic applications.. Certificate ISO ...

Considering the crucial role of miRNAs in vascular remodeling, therapeutic strategies to either increase or inhibit miRNAs might be a promising approach. When compared with other molecular targets, the targeting of miRNAs has several theoretical advantages: 1) the antisense oligonucleotides to inhibit endogenous miRNAs are short, which makes packaging and delivery easier; 2) cell-specific expression of miRNAs and suppression of disease-specific targets may attenuate off-target effects of the treatment; and 3) because of regulatory effects of miRNAs, therapeutic modulation of miRNA levels may allow more rapid and subtle interference with molecular disease mechanisms.. Several methods for miRNA loss-of-function in vivo studies using modified antisense oligonucleotides have been developed. Intravenous administration of antagomirs, a class of nucleotides chemically modified with 2′-O-methyl phosphoramidites, and a hydroxyprolinol-linked cholesterol solid support, suppresses the expression of ...

Fig. 6 A-H: Heat shock-induced Emx3 protein partially rescues emx3 morpholino antisense oligonucleotides (MO) -induced knockdown, whereas excessive Emx3 dorsalizes the telencephalon. A-C,E-G: Low levels of Emx3-myc-GFP expression (e3-GFP+), induced by heat shock at the 1- or 2-somite stages (10.5 hr, hs2s), 20-30 min after the onset of endogenous emx3 mRNA expression at bud stage, rescues the emx3MO-induced down regulation of tcf4l (A-C) and eomesa (eom., E-G). Later heat shock, between the 5- and 7-somite stages (11.5 hr, hs5s), does not rescue the phenotype (B,D). Higher levels of GFP expression after heat shock (e3-GFP++), indicating higher levels of Emx3 activity, lead to dorsalized telencephalon as assessed by ventrally expanded expression of tcf4l (A,H). A-D,H: Frontal views, dorsal to the top, 25 hr; (E-G) Dorsal view, rostral to the left, 12-somite stage (15 hr). Scale bar = 100 μm.. ...

Your bioshots for the week: 1. WAVE Life Sciences: FDA approves orphan drug designation for its lead candidate against Huntingtons disease WAVE produces stereopure nucleic acid drugs (antisense oligos, ASOs) which reportedly improves their pharmacology. ASOs are commonly modified with phosphorothioate that improves their stability in vivo but this also introduces a new chiral center. CEO Paul Bolno (former Head of…

This study investigated the effect of agmatine (Agm) in proliferation of ovine trophecdoderm cells (oTr1) as well as the importance of the arginine decarboxylase (ADC) and agmatinase (AGMAT) alternative pathway for synthesis of polyamines in ovine conceptuses during the peri-implantation period of pregnancy. Morpholino antisense oligonucleotides (MAOs) were used to inhibit translation of mRNAs for ODC1 alone, AGMAT alone, and their combination. Rambouillet ewes (N = 50) were assigned randomly to the following treatments on Day 8 of pregnancy: MAO control (n = 10); MAO-ODC1 (n = 8); MAO-ADC (n = 6); MAO-ODC1:MAO-ADC (n = 9); or MAO-ODC1:MAO-AGMAT (n = 9 ...

Her beskriver vi en metode til at levere medicin til rat centralnervesystemet ved at implantering af et kateter i lænde intrathecale...

Cardiac sicknesses are primarily probably the most frequent causes of demise in industrialized nations. Pathological transforming of the heart muscle is introduced on by a quantity of etiologies equivalent to prolonged hypertension or accidents that will lead to myocardial infarction and in essential situations moreover the demise of the affected particular person. The micro-RNA miR-132 … Read more. ...

Previously we have shown that the expression of RA-A47 (rheumatoid arthritis-related antigen) which is identical to HSP47, a collagen-binding chaperon

Poly(amidoamine) dendrimers are the most recognized class of dendrimer. Amino-terminated (PAMAM-NH2) and hydroxyl-terminated (PAMAM-OH) dendrimers of generation 4 are widely used, since they are...

TY - JOUR. T1 - Oligonucleotides as a tool for intracellular DDS. AU - Sasaki, Shigeki. PY - 2002. Y1 - 2002. N2 - Oligonucleotides attaching a reactive molecule may cause irreversible change to the target sequence at the reactive site. Inhibition of transcription, site-directed mutagenesis (or site-directed manipulation of genes) at the specific site has been proposed as a new tool of biotechnology with the use of a triplex-forming oligonucleotide. Thus, the oligonucleotides are regarded as a tool for intracellular DOS of useful reactive molecules. This review deals with development of a new alkylating group with high selectivity toward cytosine as well as new non-natural base analogs for the formation of stable non-natural triplexes.. AB - Oligonucleotides attaching a reactive molecule may cause irreversible change to the target sequence at the reactive site. Inhibition of transcription, site-directed mutagenesis (or site-directed manipulation of genes) at the specific site has been proposed ...

Journal of Drug Discovery, Development and Delivery is an open access journal dedicated to publish articles in all areas of Drug Discovery, Development and Delivery.

MINNEAPOLIS, July 20, 2020 /PRNewswire/ -- Bio-Techne Corporation (NASDAQ:TECH) today announced the expansion of the RNAscope™ platform with the release of the miRNAscope™ Assay. The RNAscope technology is an advanced in situ hybridization assay for the spatial visualization of single-molecule RNA with single-cell resolution directly in intact tissues.. The miRNAscope Assay extends the RNAscope technology to enable the in situ detection of short nucleic acid targets between 17-50 nucleotides which includes an important class of small non-coding RNAs called microRNAs as well as short synthetic oligonucleotide therapeutics such as small interfering RNAs (siRNAs) and antisense oligonucleotides (ASOs). The assay addresses a critical need to reliably detect these ultra-short molecules in native tissue with minimal time and effort, delivering data in an easy to interpret format. Traditional assays such as microarrays, PCRs, and sequencing methods provide useful molecular profiles, but their use ...

Less than a year ago, Eli Lilly and Co. and Isis Pharmaceuticals Inc. signed a deal for Isis antisense technology worth as much as $400 million. On Tuesday the companies expanded that deal, opening it up to include specific gene targets for cancer and adding to its total value. (BioWorld Today)