A5073359920- RNA Interference and Gene Delivery
- Genomics and Chromatin Dynamics
- Advanced biosensing and bioanalysis techniques
- Avian ecology and behavior
- Physiological and biochemical adaptations
- RNA Research and Splicing
- Gene Regulatory Network Analysis
- RNA and protein synthesis mechanisms
- MicroRNA in disease regulation
- Lipid Membrane Structure and Behavior
- Insect Resistance and Genetics
- Porphyrin and Phthalocyanine Chemistry
- Fish Ecology and Management Studies
- DNA and Nucleic Acid Chemistry
- RNA modifications and cancer
- Animal Behavior and Reproduction
- Nonlinear Optical Materials Research
- Fungal and yeast genetics research
- Genetically Modified Organisms Research
- Nuclear Structure and Function
- Bat Biology and Ecology Studies
- Spectroscopy and Quantum Chemical Studies
- CRISPR and Genetic Engineering
- Molecular spectroscopy and chirality
- Plant and animal studies
Alnylam Pharmaceuticals (United States)
2016-2025
University of California, Santa Cruz
2010-2024
Bayer (United States)
2023
Dimagi (United States)
2022
California Lutheran University
2021
Monsanto (United States)
2001-2018
Sangamo BioSciences (United States)
2017
University of Regensburg
2013
Duke University
2009
Harvard University
2004-2008
Significant progress has been made in the advancement of RNAi therapeutics by combining a synthetic triantennary N-acetylgalactosamine ligand targeting asialoglycoprotein receptor with chemically modified small interfering RNA (siRNA) designs, including recently described Enhanced Stabilization Chemistry. This strategy demonstrated robust RNAi-mediated gene silencing liver after subcutaneous administration across species, human. Here we demonstrate that substantial efficacy improvements can...
Covalent attachment of a synthetic triantennary N-acetylagalactosamine (GalNAc) ligand to chemically modified siRNA has enabled asialoglycoprotein (ASGPR)-mediated targeted delivery therapeutically active siRNAs hepatocytes in vivo. This approach become transformative for the RNAi therapeutics as well other classes investigational oligonucleotide liver. For efficient functional intact drug into desired subcellular compartment, however, it is critical that nucleic acids are stabilized against...
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...
Although the spatial location of genes within nucleus has been implicated in their transcriptional status, little is known about dynamics gene that accompany large-scale changes expression. The mating haploid yeast Saccharomyces cerevisiae accompanied by a change transcription and developmental program. We examined nuclear organization stimulus pheromone α factor found most α-factor-induced become associated with components envelope. myosin-like protein Mlp1, which mRNA export, was further...
The nuclear localization of genes is intimately tied to their transcriptional status in Saccharomyces cerevisiae , with populations both active and silent interacting components the envelope. We investigated relationship between mammalian pore human genome by generating high-resolution, chromosome-wide binding maps nucleoporin 93 (Nup93) presence absence a potent histone deacetylase inhibitor (HDACI). Here, we report extensive genomic reorganization respect following HDACI treatment,...
Electron microscopy of single gene molecules and mathematical modeling shows that a promoter stochastically transitions between transcriptionally favorable unfavorable nucleosome configurations, providing mechanism for transcriptional bursting.
Abstract Preclinical mechanistic studies have pointed towards RNA interference-mediated off-target effects as a major driver of hepatotoxicity for GalNAc–siRNA conjugates. Here, we demonstrate that single glycol nucleic acid or 2′–5′-RNA modification can substantially reduce small interfering (siRNA) seed-mediated binding to transcripts while maintaining on-target activity. In siRNAs with established driven by effects, these novel designs seed-pairing destabilization, termed enhanced...
Abstract Enzyme‐linked immunosorbent assays (ELISA) and bioassays were used to estimate levels of Cry1Ab protein in four species phytophagous insects after feeding on transgenic Bt‐corn plants expressing or artificial diets containing protein. The level sources the was uniformly low but varied with insect as well food source. For corn leaf aphid, Rhopalosiphum maidis (Fitch), diet solutions protein, aphid 250–500 times less than original diet, whereas no detected by ELISA aphids Bt‐Corn...
Efficient gene silencing by RNA interference (RNAi) in vivo requires the recognition and binding of 5΄- phosphate guide strand an siRNA Argonaute protein. However, for exogenous siRNAs it is limited rapid removal metabolic enzymes. Here, we have determined crystal structure human Argonaute-2 complex with metabolically stable 5΄-(E)-vinylphosphonate (5΄-E-VP) at 2.5-Å resolution. The demonstrates how 5΄ site Mid domain able to adjust key residues 5΄-nucleotide pocket compensate change...
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...
Chromatin comprises nucleosomes as well nonnucleosomal histone–DNA particles. Prenucleosomes are rapidly formed particles that can be converted into canonical by a motor protein such ACF. Here we show the prenucleosome is stable conformational isomer of nucleosome. It consists histone octamer associated with ∼80 base pair (bp) DNA, which located at position corresponds to central 80 bp nucleosome core particle. Monomeric prenucleosomes free flanking DNA do not spontaneously fold but an...
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...
The role of the actin cytoskeleton in regulating membrane protein trafficking is complex and depends on cell type being examined. Using epididymis as a model system which luminal acidification crucial for sperm maturation storage, we now report that modulation by calcium-activated actin-capping -severing gelsolin plays key vacuolar H(+)-ATPase (V-ATPase) recycling. Epididymal clear cells contain abundant V-ATPase their apical pole, an increase cell-surface expression correlates with proton...
We measured carbon dioxide (CO₂) production and water influx rates of free-ranging, foraging, wandering albatrosses (Diomedea exulans) attending chicks, by means doubly labeled water. CO₂ for nine birds averaged 0.656 ± 0.079 ml g⁻¹ h⁻¹. This is equivalent to an overall energy expenditure 3,354 kJ bird⁻¹ day⁻¹ or 1.83 times basal metabolic rate (BMR). the lowest field recorded free-ranging attributable relatively low cost soaring flight used as opposed flapping flight. Energy was estimated...
Abstract Fluctuations in the initiation rate of transcription, first step gene expression, ensue from stochastic behavior molecular process that controls transcription. In steady state, regulatory is often assumed to operate reversibly, i.e., equilibrium. However, reversibility imposes fundamental limits information processing. For instance, assumption equilibrium difficult square with precision which executes its task eukaryotes. Here we provide evidence — microscopic analyses transcription...
The conjugation of the sense strands small interfering RNA (siRNA) to tri-N-acetylgalactosamine (GalNAc), ligand for a hepatocyte-specific receptor, enables delivery multiple clinically approved therapeutic agents that act through interference pathway. Here, we report systematic evaluation siRNAs with 3′ termini antisense conjugated GalNAc first time. These designs retained same receptor affinity, in vitro and vivo activities, as well level loading into RNA-induced silencing complex...
The purpose of this study was to assess the persistence and accumulation Cry1Ab protein in soil as a result sustained planting genetically modified Bacillus thuringiensis (Bt) corn hybrids. Soil samples were collected from agricultural fields five corn-growing regions United States where Bt hybrids (MON 810 or Bt11) had been planted for at least 3 consecutive yr. At each site, during period (postanthesis) again within 6 wk after harvest. Multiple specimens matched cornfields nearby, non-Bt...
The diets of adult Macaroni penguins Eudyptes chrysolophus and Southern rockhopper E. chrysocome were analysed quantitatively at Marion Island, southern Indian Ocean, throughout two successive chick‐rearing seasons. broadly similar. Crustaceans the predominant prey type comprising, overall, 90% by mass 98% numbers in 96% 99% penguins. Nauticaris marionis was crustacean eaten both penguin species 1983–84, but Euphausia vallentini Thysanoessa vicina predominated 1984–85. Themisto gaudichaudii...
(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...