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Challenges of Assessing Exon 53 Skipping of the Human DMD Transcript with Locked Nucleic Acid-Modified Antisense Oligonucleotides in a Mouse Model for Duchenne Muscular Dystrophy. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-12-01 Sarah Engelbeen,Daniel O'Reilly,Davy Van De Vijver,Ingrid Verhaart,Maaike van Putten,Vignesh Hariharan,Matthew Hassler,Anastasia Khvorova,Masad J Damha,Annemieke Aartsma-Rus
Antisense oligonucleotide (AON)-mediated exon skipping is a promising therapeutic approach for Duchenne muscular dystrophy (DMD) patients to restore dystrophin expression by reframing the disrupted open reading frame of the DMD transcript. However, the treatment efficacy of the already conditionally approved AONs remains low. Aiming to optimize AON efficiency, we assessed exon 53 skipping of the DMD
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Impact of the Inhibition of Organic Anion Transporter on Tricyclo-DNA-Mediated Exon Skipping in the mdx Mouse Model. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-11-15 Flavien Bizot,Thomas Tensorer,Luis Garcia,Aurélie Goyenvalle
Antisense-mediated exon skipping is one of the most promising therapeutic strategies for Duchenne muscular dystrophy (DMD) and some antisense oligonucleotide (ASO) drugs have already been approved by the U.S. FDA for DMD. The potential of this therapy is still limited by several challenges including the poor distribution of ASOs to target tissues. Indeed, most of them accumulate in the kidney and tend
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Cholesterol-Conjugated Supramolecular Multimeric siRNAs: Effect of siRNA Length on Accumulation and Silencing In Vitro and In Vivo. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-11-09 Ivan V Chernikov,Ul'yana A Ponomareva,Mariya I Meschaninova,Irina K Bachkova,Anna A Teterina,Daniil V Gladkikh,Innokenty A Savin,Valentin V Vlassov,Marina A Zenkova,Elena L Chernolovskaya
Conjugation of small interfering RNA (siRNA) with lipophilic molecules is one of the most promising approaches for delivering siRNA in vivo. The rate of molecular weight-dependent siRNA renal clearance is critical for the efficiency of this process. In this study, we prepared cholesterol-containing supramolecular complexes containing from three to eight antisense strands and examined their accumulation
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Development of Long Asymmetric siRNA Structure for Target Gene Silencing and Immune Stimulation in Mammalian Cells. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-10-01 Soonkap Kim,Young Gyu Kang,Jaejin Kim,Pooja Dua,Dong-Ki Lee
Post-transcriptional regulation of transcript abundances by RNA interference (RNAi) is a widely conserved regulatory mechanism to control cellular processes. We previously introduced an alternative siRNA structure called asymmetric siRNA (asiRNA), and showed that asiRNA exhibits comparable gene-silencing efficiency with reduced off-target effects compared with conventional siRNAs. However, to what
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Highly Potent Antisense Oligonucleotides Locked Nucleic Acid Gapmers Targeting the SARS-CoV-2 RNA Genome. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-09-29 Vita Dauksaite,Ali Tas,Falk Wachowius,Anouk Spruit,Martijn J van Hemert,Eric J Snijder,Eric P van der Veer,Anton Jan van Zonneveld
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused the current worldwide pandemic and the associated coronavirus disease 2019 with potentially lethal outcome. Although effective vaccines strongly contributed to reduce disease severity, establishing a toolbox to control current and newly emerging coronaviruses of epidemic concern requires the development of novel therapeutic
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Delivery Characterization of SPL84 Inhaled Antisense Oligonucleotide Drug for 3849 + 10 kb C- > T Cystic Fibrosis Patients. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-08-29 Efrat Ozeri-Galai,Lital Friedman,Ofra Barchad-Avitzur,Matthew R Markovetz,William Boone,Kaitlyn R Rouillard,Chava D Stampfer,Yifat S Oren,David B Hill,Batsheva Kerem,Gili Hart
Recent advances in the therapeutic potential of RNA-related treatments, specifically for antisense oligonucleotide (ASO)-based drugs, have led to increased numbers of ASO regulatory approvals. In this study, we focus on SPL84, an inhaled ASO-based drug, developed for the treatment of the pulmonary disease cystic fibrosis (CF). Pulmonary drug delivery is challenging, due to a variety of biological,
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New Oligonucleotide 2'-O-Alkyl N3'→P5' (Thio)-Phosphoramidates as Potent Antisense Agents: Physicochemical Properties and Biological Activity. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-08-28 Saúl Martínez-Montero,Vivek K Rajwanshi,Rajendra K Pandey,N Tilani S De Costa,Jin Hong,Leonid Beigelman,Sergei M Gryaznov,Soheil Pourshahian
We describe here the design, synthesis, physicochemical properties, and hepatitis B antiviral activity of new 2'-O-alkyl ribonucleotide N3'→P5' phosphoramidate (2'-O-alkyl-NPO) and (thio)-phosphoramidite (2'-O-alkyl-NPS) oligonucleotide analogs. Oligonucleotides with different 2'-O-alkyl modifications such as 2'-O-methyl, -O-ethyl, -O-allyl, and -O-methoxyethyl combined with 3'-amino sugar-phosphate
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OSWG Recommended Approaches to the Nonclinical Pharmacokinetic (ADME) Characterization of Therapeutic Oligonucleotides. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-08-17 Cindy L Berman,Madeleine Antonsson,Sandor Batkai,Sieto Bosgra,Girish R Chopda,Wouter Driessen,Jeffrey Foy,Chopie Hassan,Xiao Shelley Hu,Hyun Gyung Jang,Meena,Mark Sanseverino,Thomas Thum,Yanfeng Wang,Martin Wild,Jing-Tao Wu
This white paper summarizes the recommendations of the absorption, distribution, metabolism, and excretion (ADME) Subcommittee of the Oligonucleotide Safety Working Group for the characterization of absorption, distribution, metabolism, and excretion of oligonucleotide (ON) therapeutics in nonclinical studies. In general, the recommended approach is similar to that for small molecule drugs. However
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From Failure to Meet the Clinical Endpoint to U.S. Food and Drug Administration Approval: 15th Antisense Oligonucleotide Therapy Approved Qalsody (Tofersen) for Treatment of SOD1 Mutated Amyotrophic Lateral Sclerosis. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-08-01 Willeke van Roon-Mom,Chantal Ferguson,Annemieke Aartsma-Rus
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mTOR Inhibition Enhances Delivery and Activity of Antisense Oligonucleotides in Uveal Melanoma Cells. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-06-30 Shanna Dewaele,Louis Delhaye,Boel De Paepe,Bram Bogaert,Ramiro Martinez,Jasper Anckaert,Nurten Yigit,Justine Nuytens,Rudy Van Coster,Sven Eyckerman,Koen Raemdonck,Pieter Mestdagh
Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Owing to a lack of effective treatments, patients with metastatic disease have a median survival time of 6-12 months. We recently demonstrated that the Survival Associated Mitochondrial Melanoma Specific Oncogenic Non-coding RNA (SAMMSON) is essential for UM cell survival and that antisense oligonucleotide (ASO)-mediated
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Sequence-Controlled Spherical Nucleic Acids: Gene Silencing, Encapsulation, and Cellular Uptake. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-05-17 Sepideh Kaviani,Hassan H Fakih,Jathavan Asohan,Adam Katolik,Masad J Damha,Hanadi F Sleiman
Antisense oligonucleotides (ASOs) can predictably alter RNA processing and control protein expression; however, challenges in the delivery of these therapeutics to specific tissues, poor cellular uptake, and endosomal escape have impeded progress in translating these agents into the clinic. Spherical nucleic acids (SNAs) are nanoparticles with a DNA external shell and a hydrophobic core that arise
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Experimental Model Systems Used in the Preclinical Development of Nucleic Acid Therapeutics. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-05-05 Haiyan Zhou,Virginia Arechavala-Gomeza,Alejandro Garanto
Preclinical evaluation of nucleic acid therapeutics (NATs) in relevant experimental model systems is essential for NAT drug development. As part of COST Action "DARTER" (Delivery of Antisense RNA ThERapeutics), a network of researchers in the field of RNA therapeutics, we have conducted a survey on the experimental model systems routinely used by our members in preclinical NAT development. The questionnaire
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Complement C3d/C4d Deposition on Platelets Correlates with 2'-O-Methoxyethyl Antisense Oligonucleotide-Induced Thrombocytopenia in Monkeys. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-04-21 Lijiang Shen,Andrea Wong,Satoru Oneda,Brian R Curtis,Joe Schroeder,Tom Zanardi,Jeffery A Engelhardt,Scott P Henry,Padmakumar Narayanan
2'-O-Methoxyethyl antisense oligonucleotide (2'-MOE ASO)-induced severe thrombocytopenia (TCP) [platelet (PLT) count <50 K/μL] was observed in the Asian-sourced cynomolgus monkeys with low incidence (2%-4% at doses >5 mg/kg/week). The potential mechanisms for TCP were studied using the Mauritian-sourced cynomolgus monkeys, which were shown to be more susceptible to ASO-induced TCP, along with the Asian-sourced
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Positive Allosteric Modulation of Antithrombin's Inhibitory Activity by RNA Aptamers. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-04-21 Khalequz Zaman,Adi Breitman,Isa Malik,Yolanda M Fortenberry
The leading cause of death in adults in the United States is cardiovascular disease, with mortality and morbidity mainly attributed to thromboembolism. Heparin is the most common therapy used for treating venous and arterial thrombosis. Heparin effectively accelerates the inhibition of coagulation proteases thrombin and factor Xa through the serine protease inhibitor (serpin) antithrombin (AT). Heparin
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DNAzymes: Expanding the Potential of Nucleic Acid Therapeutics. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-04-21 Leon M Larcher,Ianthe L Pitout,Niall P Keegan,Rakesh N Veedu,Sue Fletcher
Nucleic acids drugs have been proven in the clinic as a powerful modality to treat inherited and acquired diseases. However, key challenges including drug stability, renal clearance, cellular uptake, and movement across biological barriers (foremost the blood-brain barrier) limit the translation and clinical efficacy of nucleic acid-based therapies, both systemically and in the central nervous system
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Next Generation Exon 51 Skipping Antisense Oligonucleotides for Duchenne Muscular Dystrophy. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-04-10 Judith van Deutekom,Chantal Beekman,Suzanne Bijl,Sieto Bosgra,Rani van den Eijnde,Dennis Franken,Bas Groenendaal,Bouchra Harquouli,Anneke Janson,Paul Koevoets,Melissa Mulder,Daan Muilwijk,Galyna Peterburgska,Bianca Querido,Janwillem Testerink,Ruurd Verheul,Peter de Visser,Rudie Weij,Annemieke Aartsma-Rus,Jukka Puoliväli,Timo Bragge,Charles O'Neill,Nicole A Datson
In the last two decades, antisense oligonucleotides (AONs) that induce corrective exon skipping have matured as promising therapies aimed at tackling the dystrophin deficiency that underlies the severe and progressive muscle fiber degeneration in Duchenne muscular dystrophy (DMD) patients. Pioneering first generation exon 51 skipping AONs like drisapersen and eteplirsen have more recently been followed
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Rapid and Reliable Quantification of Prime Editing Targeting Within the Porcine ABCA4 Gene Using a BRET-Based Sensor. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-03-01 Tobias Wimmer,Hannah Sawinski,Anne M Urban,Jan Motlik,Knut Stieger
Stargardt disease (STGD) leads to blindness in children and young adults. So far, no curative therapy is available and gene augmentation therapies have not yet advanced to the clinics, in part, due to the limited packaging capacity of adeno-associated viruses used to transfer genes into photoreceptor cells. Prime editing offers a new perspective to treat mutations on the genomic level. A nicking variant
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Aerosolized Pulmonary Delivery of mRNA Constructs Attenuates Severity of Escherichia coli Pneumonia in the Rat. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-22 Sean D McCarthy,Christopher B Rohde,Matt Angel,Claire H Masterson,Ronan MacLoughlin,Juan Fandiño,Héctor E González,Declan Byrnes,John G Laffey,Daniel O'Toole
Acute respiratory distress syndrome (ARDS), a rapid onset inflammatory lung disease with no effective specific therapy, typically has pathogenic etiology termed pneumonia. In previous studies nuclear factor-κB (NF-κB) inhibitor α super-repressor (IκBα-SR) and extracellular superoxide dismutase 3 (SOD3) reduced pneumonia severity when prophylactically delivered by viral vector. In this study, mRNA coding
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Considerations for the Terminal Sterilization of Oligonucleotide Drug Products. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-14 Daniel Paul DeCollibus,Justin Searcy,Anna Tivesten,Nadim Akhtar,Christian Lindenberg,Nounja Abarrou,Sujana Pradhan,Maggie Fiandaca,Jenny Franklin,Geetha Govindan,Hung-Yi Liu,David Royle,Patrick Lim Soo,Kirsten Storch
A primary function of the parenteral drug product manufacturing process is to ensure sterility of the final product. The two most common methods for sterilizing parenteral drug products are terminal sterilization (TS), whereby the drug product is sterilized in the final container following filling and finish, and membrane sterilization, whereby the product stream is sterilized by membrane filtration
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Systematic Analysis of Chemical Modifications of Phosphorothioate Antisense Oligonucleotides that Modulate Their Innate Immune Response. Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-07 Adam J Pollak,Luyi Zhao,Stanley T Crooke
While rare, some gapmer phosphorothioate (PS) antisense oligonucleotides (ASOs) can induce a noncanonical TLR9-dependent innate immune response. In this study, we performed systematic analyses of the roles of PS ASO backbone chemistry, 2' modifications, and sequence in PS ASO induced TLR9 signaling. We found that each of these factors can contribute to altering PS ASO induced TLR9 signaling, and in
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Chemical Modifications and Design Influence the Potency of Huntingtin Antigene Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-03 Osama Saher, Eman M. Zaghloul, Tea Umek, Daniel W. Hagey, Negin Mozafari, Mathias B. Danielsen, Alaa S. Gouda, Karin E. Lundin, Per T. Jørgensen, Jesper Wengel, C.I. Edvard Smith, Rula Zain
Huntington's disease is a neurodegenerative, trinucleotide repeat (TNR) disorder affecting both males and females. It is caused by an abnormal increase in the length of CAG•CTG TNR in exon 1 of the Huntingtin gene (HTT). The resultant, mutant HTT mRNA and protein cause neuronal toxicity, suggesting that reduction of their levels would constitute a promising therapeutic approach. We previously reported
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Drug Metabolism and Pharmacokinetics of Antisense Oligonucleotide Therapeutics: Typical Profiles, Evaluation Approaches, and Points to Consider Compared with Small Molecule Drugs Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-03 Hideo Takakusa, Norihiko Iwazaki, Makiya Nishikawa, Tokuyuki Yoshida, Satoshi Obika, Takao Inoue
Oligonucleotide therapeutics are attracting attention as a new treatment modality for a range of diseases that have been difficult to target using conventional approaches. Technical advances in chemical modification and drug delivery systems have led to the generation of compounds with excellent profiles as pharmaceuticals, and 16 oligonucleotide therapeutics have been marketed to date. There is a
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Considerations in the Preclinical Assessment of the Safety of Antisense Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Aurélie Goyenvalle, Cecilia Jimenez-Mallebrera, Willeke van Roon, Sabine Sewing, Arthur M. Krieg, Virginia Arechavala-Gomeza, Patrik Andersson
The nucleic acid therapeutics field has made tremendous progress in the past decades. Continuous advances in chemistry and design have led to many successful clinical applications, eliciting even more interest from researchers including both academic groups and drug development companies. Many preclinical studies in the field focus on improving the delivery of antisense oligonucleotide drugs (ONDs)
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Consensus Guidelines for the Design and In Vitro Preclinical Efficacy Testing N-of-1 Exon Skipping Antisense Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Annemieke Aartsma-Rus, Alejandro Garanto, Willeke van Roon-Mom, Erin M. McConnell, Victoria Suslovitch, Winston X. Yan, Jonathan K. Watts, Timothy W. Yu
Antisense oligonucleotides (ASOs) can modulate pre-mRNA splicing. This offers therapeutic opportunities for numerous genetic diseases, often in a mutation-specific and sometimes even individual-specific manner. Developing therapeutic ASOs for as few as even a single patient has been shown feasible with the development of Milasen for an individual with Batten disease. Efforts to develop individualized
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Efficient Downregulation of Alk4 in Skeletal Muscle After Systemic Treatment with Conjugated siRNAs in a Mouse Model for Duchenne Muscular Dystrophy Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Sarah Engelbeen, Svetlana Pasteuning-Vuhman, Joke Boertje-van der Meulen, Rubina Parmar, Klaus Charisse, Laura Sepp-Lorenzino, Muthiah Manoharan, Annemieke Aartsma-Rus, Maaike van Putten
Downregulation of genes involved in the secondary pathology of Duchenne muscular dystrophy, for example, inflammation, fibrosis, and adiposis, is an interesting approach to ameliorate degeneration of muscle and replacement by fibrotic and adiposis tissue. Small interfering RNAs (siRNAs) are able to downregulate target genes, however, delivery of siRNAs to skeletal muscle still remains a challenge.
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Cholesterol-Conjugated siRNA Silencing Tnf for the Treatment of Liver Macrophage-Mediated Acute Inflammation in Nonalcoholic Fatty Liver Disease Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Kevin Craig, Marc Abrams, Mansoor Amiji
Despite wide recognition as a disease of pandemic proportions, effective treatments for nonalcoholic fatty liver disease (NAFLD) remain elusive. Most of the current clinical programs aim to reduce hepatic fat accumulation and, thus, prevent downstream inflammation and fibrosis. To date, this therapeutic approach has helped identify a potential disconnect between steatosis reduction and disease resolution
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Knockdown of Circular RNAs Using LNA-Modified Antisense Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Marianne Bengtson Løvendorf, Anja Holm, Andreas Petri, Charlotte Albæk Thrue, Shizuka Uchida, Morten T. Venø, Sakari Kauppinen
Circular RNAs (circRNAs) constitute an abundant class of covalently closed noncoding RNA molecules that are formed by backsplicing from eukaryotic protein-coding genes. Recent studies have shown that circRNAs can act as microRNA or protein decoys, as well as transcriptional regulators. However, the functions of most circRNAs are still poorly understood. Because circRNA sequences overlap with their
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Elevation of SHANK3 Levels by Antisense Oligonucleotides Directed Against the 3′-UTR of the Human SHANK3 mRNA Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Nadine Stirmlinger, Jan Philip Delling, Stefanie Pfänder, Tobias M. Boeckers
SHANK3 is a member of the SHANK family of scaffolding proteins that localize to the postsynaptic density of excitatory synapses. Mutations within the SHANK3 gene or SHANK3 haploinsufficiency is thought to be one of the major causes for Phelan-McDermid Syndrome (PMDS) that is characterized by a broad spectrum of autism-related behavioral alterations. Several approaches have already been proposed to
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Integrated Assessment of Phase 2 Data on GalNAc3-Conjugated 2′-O-Methoxyethyl-Modified Antisense Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2023-02-01 Brenda F. Baker, Shuting Xia, Wesley Partridge, T. Jesse Kwoh, Sotirios Tsimikas, Sanjay Bhanot, Richard S. Geary
Receptor-mediated delivery of an antisense oligonucleotide (ASO) using the ligand-conjugated antisense technology is establishing a new benchmark for antisense therapeutics. The triantennary N-acetylgalactosamine (GalNAc3) cluster is the first conjugated ligand to yield a marked increase in ASO potency for RNA targets expressed by hepatocytes, compared to the unconjugated form. In this study, we present
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Translational Pharmacokinetic/Pharmacodynamic Model for mRNA-3927, an Investigational Therapeutic for the Treatment of Propionic Acidemia Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-27 Husain Attarwala, Matthew Lumley, Min Liang, Vijay Ivaturi, Joe Senn
Propionic acidemia (PA) is an ultrarare disorder caused by deficiency of the mitochondrial enzyme, propionyl-CoA carboxylase (PCC), composed of PCCA and PCCB subunits. An enzyme replacement therapy is being developed using dual messenger RNA (mRNA) therapy composed of lipid nanoparticles (LNPs) encapsulating mRNAs encoding PCCA and PCCB subunits of the PCC enzyme. We herein report on development of
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Time Is a Critical Factor When Evaluating Oligonucleotide Therapeutics in Human Ether-a-Go-Go-Related Gene Assays Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-26 Yusheng Qu, Robert Kirby, Richard Davies, Ayesha Jinat, Stefano Stabilini, Bin Wu, Longchuan Yu, BaoXi Gao, Hugo M. Vargas
In accord with International Conference on Harmonization S7B guidelines, an in vitro human ether-a-go-go-related gene (hERG) assay is one component of an integrated risk assessment for delayed ventricular repolarization. Function of hERG could be affected by direct (acute) mechanisms, or by indirect (chronic) mechanisms. Some approved oligonucleotide therapeutics had submitted hERG data to regulatory
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SIDT2 Inhibits Phosphorothioate Antisense Oligonucleotide Activity by Regulating Cellular Localization of Lysosomes Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-19 Jing Crystal Zhao, Aurian Saleh, Stanley T. Crooke
Phosphorothioate (PS)-modified antisense oligonucleotide (ASO) drugs enter cells through endocytic pathways where a majority are entrapped within membrane-bound endosomes and lysosomes, representing a limiting step for antisense activity. While late endosomes have been identified as a major site for productive PS-ASO release, how lysosomes regulate PS-ASO activity beyond macromolecule degradation remains
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The Quest for mRNA Vaccines Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Eli Gilboa, David Boczkowski, Smita K. Nair
The success of mRNA vaccines against COVID-19 is nothing short of a medical revolution. Given its chemical lability the use of mRNA as a therapeutic has been counterintuitive and met with skepticism. The development of mRNA-based COVID-19 vaccines was the culmination of long and painstaking efforts by many investigators spanning over 30 years and culminating with the seminal studies of Kariko and Weissman
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Early-Stage Identification and Avoidance of Antisense Oligonucleotides Causing Species-Specific Inflammatory Responses in Human Volunteer Peripheral Blood Mononuclear Cells Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Sebastien A. Burel, Todd Machemer, Brenda F. Baker, T. Jesse Kwoh, Suzanne Paz, Husam Younis, Scott P. Henry
A human peripheral blood mononuclear cell (PBMC)-based assay was developed to identify antisense oligonucleotide (ASO) with the potential to activate a cellular innate immune response outside of an acceptable level. The development of this assay was initiated when ISIS 353512 targeting the messenger ribonucleic acid for human C-reactive protein (CRP) was tested in a phase I clinical trial, in which
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Inflammatory Non-CpG Antisense Oligonucleotides Are Signaling Through TLR9 in Human Burkitt Lymphoma B Bjab Cells Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Adam J. Pollak, Patrick Cauntay, Todd Machemer, Suzanne Paz, Sagar Damle, Scott P. Henry, Sebastien A. Burel
Nucleic acid-based phosphorothioate containing antisense oligonucleotides (PS-ASOs) have the potential to activate cellular innate immune responses, and the level of activation can vary quite dramatically with sequence. Minimizing the degree of proinflammatory effect is one of the main selection criteria for compounds intended to move into clinical trials. While a recently developed human peripheral
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Enhancing SIRT1 Gene Expression Using Small Activating RNAs: A Novel Approach for Reversing Metabolic Syndrome Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Pinelopi Andrikakou, Vikash Reebye, Daniel Vasconcelos, Sorah Yoon, Jon Voutila, Andrew J.T. George, Piotr Swiderski, Robert Habib, Matthew Catley, David Blakey, Nagy A. Habib, John J. Rossi, Kai-Wen Huang
Metabolic syndrome (MetS) is a pathological condition characterized by abdominal obesity, insulin resistance, hypertension, and hyperlipidemia. Sirtuin 1 (SIRT1), a highly conserved histone deacetylase, is characterized as a key metabolic regulator and protector against aging-associated pathologies, including MetS. In this study, we investigate the therapeutic potential of activating SIRT1 using small
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Nucleic Acid Aptamers Increase the Anticancer Efficiency and Reduce the Toxicity of Cisplatin-Arabinogalactan Conjugates In Vivo Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Tatiana N. Zamay, Alexander K. Starkov, Olga S. Kolovskaya, Galina S. Zamay, Dmitry V. Veprintsev, Natalia Luzan, Elena D. Nikolaeva, Kirill A. Lukyanenko, Polina V. Artyushenko, Irina A. Shchugoreva, Yury E. Glazyrin, Anastasia A. Koshmanova, Alexey V. Krat, Dariya S. Tereshina, Sergey S. Zamay, Yuriy S. Pats, Ruslan A. Zukov, Felix N. Tomilin, Maxim V. Berezovski, Anna S. Kichkailo
Cisplatin is an effective drug for treating various cancer types. However, it is highly toxic for both healthy and tumor cells. Therefore, there is a need to reduce its therapeutic dose and increase targeted bioavailability. One of the ways to achieve this could be the coating of cisplatin with polysaccharides and specific carriers for targeted delivery. Nucleic acid aptamers could be used as carriers
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Cross-Species Translation of Biophase Half-Life and Potency of GalNAc-Conjugated siRNAs Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-12-05 Alessandro Boianelli, Yasunori Aoki, Maxim Ivanov, Anders Dahlén, Peter Gennemark
Small interfering RNAs (siRNAs) with N-acetylgalactosamine (GalNAc) conjugation for improved liver uptake represent an emerging class of drugs to treat liver diseases. Understanding how pharmacokinetics and pharmacodynamics translate is pivotal for in vivo study design and human dose prediction. However, the literature is sparse on translational data for this modality, and pharmacokinetics in the liver
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Assessment of the Immunogenicity Potential for Oligonucleotide-Based Drugs Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Scott P. Henry, Cecilia Arfvidsson, Josh Arrington, Jasna Canadi, Dave Crowe, Shalini Gupta, Sabine Lohmann, Benoit Massonnet, Daniel Mytych, Tina Rogers, Hobart Rogers, Chris Stebbins, Craig Stovold, Daniela Verthelyi, Adam Vigil, Chi Xuan, Yuanxin Xu, Rosie Yu, Thomas Klem
Therapeutic oligonucleotides (ONs) have characteristics of both small molecules and biologics. Although safety assessment of ONs largely follows guidelines established for small molecules, the unique characteristics of ONs often require incorporation of concepts from the safety assessment of biologics. The assessment of immunogenicity for ON therapeutics is one area where the approach is distinct from
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Host-Directed Targeting of LincRNA-MIR99AHG Suppresses Intracellular Growth of Mycobacterium tuberculosis Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Lorna Gcanga, Ousman Tamgue, Mumin Ozturk, Shandre Pillay, Raygaana Jacobs, Julius Ebua Chia, Stanley Kimbung Mbandi, Malika Davids, Keertan Dheda, Sebastian Schmeier, Tanvir Alam, Sugata Roy, Harukazu Suzuki, Frank Brombacher, Reto Guler
Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) kills 1.6 million people worldwide every year, and there is an urgent need for targeting host–pathogen interactions as a strategy to reduce mycobacterial resistance to current antimicrobials. Noncoding RNAs are emerging as important regulators of numerous biological processes and avenues for exploitation in host-directed therapeutics. Although
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Inhibition of Epidermal Growth Factor Receptor Signaling by Antisense Oligonucleotides as a Novel Approach to Epidermal Growth Factor Receptor Inhibition Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Thushara W. Madanayake, Eric A. Welsh, Lancia N.F. Darville, John M. Koomen, Charles E. Chalfant, Eric B. Haura, Timothy J. Robinson
We report a novel method to inhibit epidermal growth factor receptor (EGFR) signaling using custom morpholino antisense oligonucleotides (ASOs) to drive expression of dominant negative mRNA isoforms of EGFR by ASO-induced exon skipping within the transmembrane (16) or tyrosine kinase domains (18 and 21). In vivo ASO formulations induced >95% exon skipping in several models of nonsmall cell lung cancer
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The Combination of Mesyl-Phosphoramidate Inter-Nucleotide Linkages and 2′-O-Methyl in Selected Positions in the Antisense Oligonucleotide Enhances the Performance of RNaseH1 Active PS-ASOs Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Lingdi Zhang, Xue-hai Liang, Cheryl Li De Hoyos, Michael Migawa, Joshua G. Nichols, Graeme Freestone, Jun Tian, Punit P. Seth, Stanley T. Crooke
Antisense oligonucleotides (ASOs) that mediate RNA target degradation by RNase H1 are used as drugs to treat various diseases. Previously we found that introduction of a single 2′-O-methyl (2′-OMe) modification in position 2 of the central deoxynucleotide region of a gapmer phosphorothioate (PS) ASO, in which several residues at the termini are 2′-methoxyethyl, 2′ constrained ethyl, or locked nucleic
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Binary Antisense Oligonucleotide Agent for Cancer Marker-Dependent Degradation of Targeted RNA Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Valeriia S. Drozd, Ahmed A. Eldeeb, Dmitry M. Kolpashchikov, Daria D. Nedorezova
Antisense oligonucleotide technology is one of the most successful gene therapy (GT) approaches. However, low selectivity of antisense agents limits their application as anticancer drugs. To achieve activation of antisense agent selectively in cancer cells, herein, we propose the concept of binary antisense oligonucleotide (biASO) agent. biASO recognizes an RNA sequence of a gene associated with cancer
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Antisense Oligonucleotide Rescue of Deep-Intronic Variants Activating Pseudoexons in the 6-Pyruvoyl-Tetrahydropterin Synthase Gene Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Ainhoa Martínez-Pizarro, Fátima Leal, Lise Lolle Holm, Thomas K. Doktor, Ulrika S.S. Petersen, María Bueno, Beat Thöny, Belén Pérez, Brage S. Andresen, Lourdes R. Desviat
We report two new 6-pyruvoyl-tetrahydropterin synthase splicing variants identified through genomic sequencing and transcript analysis in a patient with tetrahydrobiopterin deficiency, presenting with hyperphenylalaninemia and monoamine neurotransmitter deficiency. Variant c.243 + 3A>G causes exon 4 skipping. The deep-intronic c.164–672C>T variant creates a potential 5′ splice site that leads to the
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Delivery of RNA Therapeutics: The Great Endosomal Escape! Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Steven F. Dowdy, Ryan L. Setten, Xian-Shu Cui, Satish G. Jadhav
RNA therapeutics, including siRNAs, antisense oligonucleotides, and other oligonucleotides, have great potential to selectively treat a multitude of human diseases, from cancer to COVID to Parkinson's disease. RNA therapeutic activity is mechanistically driven by Watson–Crick base pairing to the target gene RNA without the requirement of prior knowledge of the protein structure, function, or cellular
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Elimination of Off-Target Effect by Chemical Modification of 5′-End of siRNA Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-10-14 Yasuo Shiohama, Ryosuke Fujita, Maika Sonokawa, Masaaki Hisano, Yojiro Kotake, Marija Krstic-Demonacos, Constantinos Demonacos, Gengo Kashiwazaki, Takashi Kitayama, Masayuki Fujii
In this study, the efficiency of RNA interference of small interfering RNAs (siRNAs) bearing 5′-O-methyl-2′-deoxythymidine (X) and 5′-amino-2′, 5′-dideoxythymidine (Z) at the 5′-end of the sense strand and the antisense strand of siRNA was investigated in HeLa cells stably expressing enhanced green fluorescent protein. The results indicated that when one strand of siRNA was modified with X or Z and
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Aptamers Targeting Hallmark Proteins of Neurodegeneration Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Niloufar Mollasalehi, Liberty Francois-Moutal, David Porciani, Donald H. Burke, May Khanna
Neurodegeneration is a progressive deterioration of neural structures leading to cognitive or motor impairment of the affected patient. There is still no effective therapy for any of the most common neurodegenerative diseases (NDs) such as Alzheimer's or Parkinson's disease. Although NDs exhibit distinct clinical characteristics, many are characterized by the accumulation of misfolded proteins or peptide
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An Integrative Multi-Omics Analysis Reveals MicroRNA-143 as a Potential Therapeutic to Attenuate Retinal Angiogenesis Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Jiang-Hui Wang, Yu-Fan Chuang, Jinying Chen, Vikrant Singh, Fan-Li Lin, Richard Wilson, Leilei Tu, Chenkai Ma, Raymond C.B. Wong, Peng-Yuan Wang, Jingxiang Zhong, Alex W. Hewitt, Peter van Wijngaarden, Gregory J. Dusting, Guei-Sheung Liu
Retinal neovascularization is a severe complication of proliferative diabetic retinopathy (PDR). MicroRNAs (miRNAs) are master regulators of gene expression that play an important role in retinal neovascularization. In this study, we show that miR-143-3p is significantly downregulated in the retina of a rat model of oxygen-induced retinopathy (OIR) by miRNA-sequencing. Intravitreal injection of synthetic
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Structural Fingerprinting of siRNA Therapeutics by Solution NMR Spectroscopy Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Owen B. Becette, Anh Tran, Jace W. Jones, John P. Marino, Robert G. Brinson
Nucleic acids are an increasingly popular platform for the development of biotherapeutics to treat a wide variety of illnesses, including diseases where traditional drug development efforts have failed. To date, there are 14 short oligonucleotide therapeutics and 2 messenger RNA (mRNA) vaccines approved by the U.S. Food and Drug Administration (FDA), which demonstrates the potential of nucleic acids
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NAT10 and DDX21 Proteins Interact with RNase H1 and Affect the Performance of Phosphorothioate Oligonucleotides Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Lingdi Zhang, Karla D. Bernardo, Timothy A. Vickers, Jun Tian, Xue-hai Liang, Stanley T. Crooke
RNase H1-dependent phosphorothioate oligonucleotides (PS-ASOs) have been developed to treat various diseases through specific degradation of target RNAs. Although many factors or features of RNA and PS-ASOs have been demonstrated to affect antisense activity of PS-ASOs, little is known regarding the roles of RNase H1-associated proteins in PS-ASO performance. In this study, we report that two nucleolar
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Targeted Delivery of Antisense Oligonucleotides Through Angiotensin Type 1 Receptor Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Carol Kuo, Mehran Nikan, Steve T. Yeh, Alfred E. Chappell, Michael Tanowitz, Punit P. Seth, Thazha P. Prakash, Adam E. Mullick
We evaluated the potential of AGTR1, the principal receptor for angiotensin II (Ang II) and a member of the G protein-coupled receptor family, for targeted delivery of antisense oligonucleotides (ASOs) in cells and tissues with abundant AGTR1 expression. Ang II peptide ASO conjugates maintained robust AGTR1 signaling and receptor internalization when ASO was placed at the N-terminus of the peptide
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Inhomogeneous Diastereomeric Composition of Mongersen Antisense Phosphorothioate Oligonucleotide Preparations and Related Pharmacological Activity Impairment Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Lorenzo Arrico, Carmine Stolfi, Irene Marafini, Giovanni Monteleone, Salvatore Demartis, Salvatore Bellinvia, Francesca Viti, Marie McNulty, Irene Cabani, Anita Falezza, Lorenzo Di Bari
Mongersen is a 21-mer antisense oligonucleotide designed to downregulate Mothers against decapentaplegic homolog 7 (SMAD7) expression to treat Crohn's disease. Mongersen was manufactured in numerous batches at different scales during several years of clinical development, which all appeared identical, using common physicochemical analytical techniques, while only phosphorous-31 nuclear magnetic resonance
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RIG-I-Mediated Innate Immune Stimulation by Chemically Synthesized Long Double-Stranded RNAs Is Structure and Sequence Dependent Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Young Gyu Kang, Jaejin Kim, Kyeongmin Lee, Jeong Yong Choe, Pooja Dua, Dong Ki Lee
Double-stranded RNAs (dsRNAs) longer than 30 bp have not been considered desirable RNA interference (RNAi) triggering structures in mammalian cells as they nonspecifically activate innate immune response. However, in earlier studies, not only dsRNA length but also 5′-triphosphate moiety produced by in vitro transcription might have affected the stimulation of innate immune system. Herein, using chemically
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Exon Skipping Through Chimeric Antisense U1 snRNAs to Correct Retinitis Pigmentosa GTPase-Regulator (RPGR) Splice Defect Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Giuseppina Covello, Gehan H. Ibrahim, Niccolò Bacchi, Simona Casarosa, Michela Alessandra Denti
Inherited retinal dystrophies are caused by mutations in more than 250 genes, each of them carrying several types of mutations that can lead to different clinical phenotypes. Mutations in Retinitis Pigmentosa GTPase-Regulator (RPGR) cause X-linked Retinitis pigmentosa (RP). A nucleotide substitution in intron 9 of RPGR causes the increase of an alternatively spliced isoform of the mature mRNA, bearing
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Improvements to Hybridization-Ligation ELISA Methods to Overcome Bioanalytical Challenges Posed by Novel Oligonucleotide Therapeutics Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-08-04 Joseph A. Haegele, Ramakrishna Boyanapalli, Jaya Goyal
As oligonucleotides (ONs) and similar nucleic acid therapeutic modalities enter development pipelines, there is continual need to develop bioanalytical methodologies addressing unique challenges they pose. Novel ONs back bone chemistries, especially those enabling stereochemical control, and base modifications are being exploited to improve pharmacological properties, potency, and increase half-lives
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Modulation of RNA Splicing by Oligonucleotides: Mechanisms of Action and Therapeutic Implications Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-06-01 Olga V. Sergeeva, Evgeniya Y. Shcherbinina, Noam Shomron, Timofei S. Zatsepin
Dysregulation of RNA splicing causes many diseases and disorders. Several therapeutic approaches have been developed to correct aberrant alternative splicing events for the treatment of cancers and hereditary diseases, including gene therapy and redirecting splicing, using small molecules or splice switching oligonucleotides (SSO). Significant advances in the chemistry and pharmacology of nucleic acid
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Combining Heparin and a FX/Xa Aptamer to Reduce Thrombin Generation in Cardiopulmonary Bypass and COVID-19 Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-06-01 Charlene V. Chabata, James W. Frederiksen, Lyra B. Olson, Ibtehaj A. Naqvi, Sharon E. Hall, Ruwan Gunaratne, Bryan D. Kraft, Loretta G. Que, Lingye Chen, Bruce A. Sullenger
Known limitations of unfractionated heparin (UFH) have encouraged the evaluation of anticoagulant aptamers as alternatives to UFH in highly procoagulant settings such as cardiopulmonary bypass (CPB). Despite progress, these efforts have not been totally successful. We take a different approach and explore whether properties of an anticoagulant aptamer can complement UFH, rather than replace it, to
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Acute Neurotoxicity of Antisense Oligonucleotides After Intracerebroventricular Injection Into Mouse Brain Can Be Predicted from Sequence Features Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-06-01 Peter H. Hagedorn, Jeffrey M. Brown, Amy Easton, Maria Pierdomenico, Kelli Jones, Richard E. Olson, Stephen E. Mercer, Dong Li, James Loy, Anja M. Høg, Marianne L. Jensen, Martin Gill, Angela M. Cacace
Antisense oligonucleotides are a relatively new therapeutic modality and safety evaluation is still a developing area of research. We have observed that some oligonucleotides can produce acute, nonhybridization dependent, neurobehavioral side effects after intracerebroventricular (ICV) dosing in mice. In this study, we use a combination of in vitro, in vivo, and bioinformatics approaches to identify
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Differential Uptake of Antisense Oligonucleotides in Mouse Hepatocytes and Macrophages Revealed by Simultaneous Two-Photon Excited Fluorescence and Coherent Raman Imaging Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-06-01 Prabuddha Mukherjee, Edita Aksamitiene, Aneesh Alex, Jindou Shi, Kajari Bera, Chi Zhang, Darold R. Spillman, Marina Marjanovic, Michael Fazio, Punit P. Seth, Kendall Frazier, Steve R. Hood, Stephen A. Boppart
Antisense oligonucleotides (ASOs), a novel paradigm in modern therapeutics, modulate cellular gene expression by binding to complementary messenger RNA (mRNA) sequences. While advances in ASO medicinal chemistry have greatly improved the efficiency of cellular uptake, selective uptake by specific cell types has been difficult to achieve. For more efficient and selective uptake, ASOs are often conjugated
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Altered Biodistribution and Hepatic Safety Profile of a Gapmer Antisense Oligonucleotide Bearing Guanidine-Bridged Nucleic Acids Nucleic Acid Ther. (IF 4.0) Pub Date : 2022-06-01 Takashi Sasaki, Yoko Hirakawa, Fumiko Yamairi, Takashi Kurita, Karin Murahashi, Hirokazu Nishimura, Norihiko Iwazaki, Hidenori Yasuhara, Takashi Tateoka, Tetsuya Ohta, Satoshi Obika, Jun Kotera
Guanidine-bridged nucleic acid (GuNA) is a novel 2′,4′-bridged nucleic acid/locked nucleic acid (2′,4′-BNA/LNA) analog containing cations that exhibit strong affinity for target RNA and superior nuclease resistance. In this study, Malat1 antisense oligonucleotide (ASO) bearing GuNA was evaluated for target knockdown (KD) activity and tolerability. The GuNA ASO did not interfere with RNase H recruitment