A5079933148- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- DNA and Nucleic Acid Chemistry
- Renin-Angiotensin System Studies
- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- HIV/AIDS drug development and treatment
- RNA and protein synthesis mechanisms
- MicroRNA in disease regulation
- Hepatitis C virus research
- Hormonal Regulation and Hypertension
- Biochemical and Molecular Research
- CRISPR and Genetic Engineering
- Carbohydrate Chemistry and Synthesis
- Virus-based gene therapy research
- RNA modifications and cancer
- Receptor Mechanisms and Signaling
- Pregnancy and preeclampsia studies
- Viral Infections and Immunology Research
- Renal and related cancers
- Click Chemistry and Applications
- Viral gastroenteritis research and epidemiology
- Alzheimer's disease research and treatments
- Neurogenesis and neuroplasticity mechanisms
- SARS-CoV-2 and COVID-19 Research
Alnylam Pharmaceuticals (United States)
2016-2025
Erasmus University Rotterdam
2024
Erasmus MC
2024
Wave Life Sciences (United States)
2017
Institut des Biomolécules Max Mousseron
2007-2011
Université de Montpellier
2006-2010
Centre National de la Recherche Scientifique
2007-2010
Laboratoire de Chimie Organique
2006
Small interfering RNAs (siRNAs) conjugated to a trivalent N-acetylgalactosamine (GalNAc) ligand are being evaluated in investigational clinical studies for variety of indications. The typical development candidate selection process includes evaluation the most active compounds toxicity rats at pharmacologically exaggerated doses. subset GalNAc-siRNAs that show rat hepatotoxicity is not advanced development. Potential mechanisms can be associated with intracellular accumulation...
One hallmark of trivalent N-acetylgalactosamine (GalNAc)-conjugated siRNAs is the remarkable durability silencing that can persist for months in preclinical species and humans. Here, we investigated underlying biology supporting this extended duration pharmacological activity. We found siRNA accumulation stability acidic intracellular compartments critical long-term show functional be liberated from these loaded into newly generated Argonaute 2 protein complexes weeks after dosing, enabling...
Homology-directed repair (HDR)–based genome editing is an approach that could permanently correct a broad range of genetic diseases. However, its utility limited by inefficient and imprecise DNA mechanisms in terminally differentiated tissues. Here, we tested Repair Drive, platform technology for selectively expanding HDR-corrected hepatocytes adult mice vivo. Drive involves transient conditioning the liver knocking down essential gene, fumarylacetoacetate hydrolase ( Fah ), delivering...
The discovery of regulatory small RNAs continues to reshape paradigms in both molecular biology and virology. Here we describe examples influenza A virus-derived viral (svRNAs). svRNAs are 22–27 nt length correspond the 5′ end each genomic RNA (vRNA) segments. Expression svRNA correlates with accumulation vRNA a bias RNA-dependent polymerase (RdRp) activity from transcription toward genome replication. Synthesis requires RdRp, nucleoprotein nuclear export protein NS2. In addition, is...
Middle East respiratory syndrome coronavirus (MERS-CoV) remains a threat to public health worldwide; however, effective vaccine or drug against CoVs unavailable. CoV helicase is one of the three evolutionary most conserved proteins in nidoviruses, thus making it an important target for development. We report here first structure full-length helicase, MERS-CoV nsp13. has multiple domains, including N-terminal Cys/His rich domain (CH) with zinc atoms, beta-barrel and C-terminal SF1 core two...
Small interfering RNAs (siRNAs) targeting hepatic angiotensinogen ( Agt ) may provide long-lasting antihypertensive effects, but the optimal approach remains unclear. Here, we assessed efficacy of a novel AGT siRNA in spontaneously hypertensive rats. Rats were treated with vehicle, (10 mg/kg fortnightly; subcutaneous), valsartan (31 per day; oral), captopril (100 valsartan+siRNA, or captopril+valsartan for 4 weeks (all groups, n=8). Mean arterial pressure (recorded via radiotelemetry) was...
BACKGROUND: Small-interfering RNA (siRNA) targeting hepatic AGT (angiotensinogen) mRNA depletes AGT, lowering blood pressure for up to 6 months. However, certain situations may require a rapid angiotensin increase. The REVERSIR (RVR) - reverse siRNA silencing technology potential approach counteract effects. METHODS: Spontaneously hypertensive rats received 10 mg/kg siRNA, and 3 weeks later were given AGT-RVR (1, 10, or 20 mg/kg). One week after dosing, redose of assessed its post-AGT-RVR...
Here we report the investigation of glycol nucleic acid (GNA), an acyclic analogue, as a modification siRNA duplexes. We evaluated impact (S)- or (R)-GNA nucleotide incorporation on RNA duplex structure by determining three individual crystal structures. These structures indicate that (S)-nucleotide backbone adopts conformation has little overall structure, while (R)-nucleotide disrupts phosphate and hydrogen bonding adjacent base pair. In addition, GNA-T nucleobase rotated in which 5-methyl...
For oligonucleotide therapeutics, chemical modifications of the sugar-phosphate backbone are frequently used to confer drug-like properties. Because 2'-deoxy-2'-fluoro (2'-F) nucleotides not known occur naturally, their safety profile was assessed when in revusiran and ALN-TTRSC02, two short interfering RNAs (siRNAs), same sequence but different modification pattern metabolic stability, conjugated an N-acetylgalactosamine (GalNAc) ligand for targeted delivery hepatocytes. Exposure...
A critical challenge for the successful development of RNA interference-based therapeutics has been enhancement their in vivo metabolic stability. In therapeutically relevant, fully chemically modified small interfering RNAs (siRNAs), modification two terminal phosphodiester linkages each strand siRNA duplex with phosphorothioate (PS) is generally sufficient to protect against exonuclease degradation vivo. Since PS are chiral, we systematically studied properties siRNAs containing single...
Chemical modifications are necessary to ensure the metabolic stability and efficacy of oligonucleotide-based therapeutics. Here, we describe analyses α-(l)-threofuranosyl nucleic acid (TNA) modification, which has a shorter 3'-2' internucleotide linkage than natural DNA RNA, in context small interfering RNAs (siRNAs). The TNA modification enhanced nuclease resistance more 2'-O-methyl or 2'-fluoro ribose modifications. TNA-containing siRNAs were prepared as triantennary N-acetylgalactosamine...
ABSTRACT Influenza A virus (IAV) is an unremitting that results in significant morbidity and mortality worldwide. Key to the viral life cycle RNA-dependent RNA polymerase (RdRp), a heterotrimeric complex responsible for both transcription replication of segmented genome. Here, we demonstrate utilizes small enhancer regulate enzymatic activity maintain stoichiometric balance We IAV synthesizes RNAs (svRNAs) interact with RdRp order promote genome segment-specific manner. svRNAs localize...
We designed novel 4′-modified 2′-deoxy-2′-fluorouridine (2′-F U) analogues with the aim to improve nuclease resistance and potency of therapeutic siRNAs by introducing 4′-C-methoxy (4′-OMe) as alpha (C4′α) or beta (C4′β) epimers. The C4′α epimer was synthesized a stereoselective route in six steps; however, both α β epimers could be obtained nonstereoselective approach starting from 2′-F U. 1H NMR analysis computational investigation α-epimer revealed that 4′-OMe imparts conformational bias...
The androgen receptor plays a critical role in the progression of prostate cancer. Here, we describe targeting prostate-specific membrane antigen using lipid nanoparticle formulation containing small interfering RNA designed to silence expression messenger encoding receptor. Specifically, Glu-urea-Lys PSMA-targeting ligand was incorporated into system formulated with long alkyl chain polyethylene glycol-lipid enhance accumulation at tumor sites and facilitate intracellular uptake cells...
Abstract Oral delivery is the most widely used and convenient route of administration medicine. However, oral hydrophilic macromolecules commonly limited by low intestinal permeability pre-systemic degradation in gastrointestinal (GI) tract. Overcoming some these challenges allowed emergence dosage forms peptide-based drugs clinical settings. Antisense oligonucleotides (ASOs) have also been investigated for but despite recent progress, bioavailability remains low. Given advancement with...
A robust, reproducible, and scalable method for the solid-phase synthesis of 5′-triphosphates DNA, RNA, their chemically modified analogues using 5′-H-phosphonate intermediates is described. 5′-Triphosphates oligonucleotides with varying lengths sequences containing different 5′-terminal nucleotides, without internal sugar-backbone modifications, were efficiently prepared as crude products or further purified by HPLC.
(E)-Vinylphosphonate ((E)-VP), a metabolically stable phosphate mimic at the 5'-end of antisense strand, enhances in vivo potency siRNA. Here we describe straightforward synthetic approach to incorporate nucleotide carrying vinylphosphonate (VP) moiety oligonucleotides under standard solid-phase synthesis and deprotection conditions by utilizing pivaloyloxymethyl (POM) protected VP-nucleoside phosphoramidites. The POM protection scope scalability 5'-VP-modified and, broader sense, modified...
Although judicious use of chemical modifications has contributed to the success nucleic acid therapeutics, poor systemic stability remains a major hurdle. The introduction functional groups around phosphate backbone can enhance nuclease resistance oligonucleotides (ONs). Here, we report synthesis enantiomerically pure (R)- and (S)-5′-C-methyl (C5′-Me) substituted nucleosides their incorporation into ONs. These generally resulted in decrease thermal oligonucleotide (ON) duplexes manner...
In this report, we investigated the hexopyranose chemical modification Altriol Nucleic Acid (ANA) within small interfering RNA (siRNA) duplexes that were otherwise fully modified with 2'-deoxy-2'-fluoro and 2'-O-methyl pentofuranose modifications. The siRNAs designed to silence transthyretin (Ttr) gene conjugated a trivalent N-acetylgalactosamine (GalNAc) ligand for targeted delivery hepatocytes. Sense antisense strands of parent duplex synthesized single ANA residues at each position on...
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