Featured
-
-
Article
| Open Access
Dynamic and static control of the off-target interactions of antisense oligonucleotides using toehold chemistry
Off-target toxicity of antisense oligonucleotides (ASOs) poses a challenge for their clinical use. Here, authors develop a toehold-bearing ASO architecture that mitigates a broad spectrum of off-target interactions, significantly enhancing the safety profile of ASO drugs.
- Chisato Terada
- , Kaho Oh
- & Tsuyoshi Yamamoto
-
Article
| Open Access
Rational design of a JAK1-selective siRNA inhibitor for the modulation of autoimmunity in the skin
Therapeutic modulation of Janus kinase family enzymes is an established approach for inflammatory and autoimmune skin diseases. Here the authors rationally design small interfering RNAs to enable single Janus kinase targeting and test this new therapeutic approach in a skin disease model for maintaining efficacy and improving selectivity.
- Qi Tang
- , Hassan H. Fakih
- & John E. Harris
-
Article
| Open Access
Adaptive design of mRNA-loaded extracellular vesicles for targeted immunotherapy of cancer
There is an emerging interest in the use of mRNA therapeutics in cancer treatment, but their precise in vivo delivery remains a challenge. Here the authors develop IFN-γ mRNA-loaded small extracellular vesicles (sEVs) with CD64 overexpressed on their surface and demonstrate its efficacy in glioblastoma mouse models resistant to immunotherapy.
- Shiyan Dong
- , Xuan Liu
- & Wen Jiang
-
Article
| Open Access
Secondary structures that regulate mRNA translation provide insights for ASO-mediated modulation of cardiac hypertrophy
The GAT4A transcription factor mediates cardiac development. Here the authors identify that the 5′ UTR of GATA4 mRNA contains a double stranded structure downstream of an upstream open reading frame (uORF) that promotes uORF-mediated suppression of the main ORF.
- Omar M. Hedaya
- , Kadiam C. Venkata Subbaiah
- & Peng Yao
-
Article
| Open Access
mRNA vaccine quality analysis using RNA sequencing
mRNA vaccines must be rigorously analysed to measure their integrity and detect contaminants, which can be time-consuming and costly. Here, authors describe a method to analyse mRNA vaccine quality using long-read sequencing and a custom bioinformatic pipeline.
- Helen M. Gunter
- , Senel Idrisoglu
- & Tim R. Mercer
-
Article
| Open Access
GalNAc-Lipid nanoparticles enable non-LDLR dependent hepatic delivery of a CRISPR base editing therapy
Kasiewicz et al. describe a structure-guided rational design approach to optimize a new GalNAc-Lipid nanoparticle that enables delivery of a base editing therapy in both low-density lipoprotein receptor-deficient and wild-type nonclinical models.
- Lisa N. Kasiewicz
- , Souvik Biswas
- & Andrew M. Bellinger
-
Article
| Open Access
Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures
Removing immunogenic uncapped mRNA from transcribed mRNA can be challenging, but is critical in mRNA research and clinical applications such as vaccines. Here, authors develop hydrophobic photocaged tag-modified cap analogs, which can be used to separate capped mRNA from uncapped mRNA, with subsequent tag removal using photo-irradiation.
- Masahito Inagaki
- , Naoko Abe
- & Hiroshi Abe
-
Article
| Open Access
Autocatalytic base editing for RNA-responsive translational control
Genetic circuits that control transgene expression in response to pre-defined transcriptional cues would enable the development of smart therapeutics. Here the authors engineer programmable RNA sensors, DART VADARs, in which ADARs autocatalytically convert target hybridization into a translational output, thus amplifying editing by endogenous ADAR via positive feedback and conferring high dynamic range and a small genetic footprint.
- Raphaël V. Gayet
- , Katherine Ilia
- & James J. Collins
-
Article
| Open Access
Single-cell quantification and dose-response of cytosolic siRNA delivery
Endosomal escape and subsequent cytosolic delivery of siRNA therapeutics is inefficient, and quantification is difficult. Here the authors report a confocal microscopy-based method to quantify cytosolic delivery of fluorescently labelled siRNA during lipid-mediated delivery.
- Hampus Hedlund
- , Hampus Du Rietz
- & Anders Wittrup
-
Article
| Open Access
Fluorinated polyamidoamine dendrimer-mediated miR-23b delivery for the treatment of experimental rheumatoid arthritis in rats
Delivery of anti-inflammatory microRNA (miRNA) could be beneficial for inflammatory diseases such as rheumatoid arthritis (RA). Here the authors show that a fluorinated polyamidoamine dendrimer nanoparticle delivers miR-23b to affected RA joints and reduces inflammation, joint damage and synovial cell influx.
- Haobo Han
- , Jiakai Xing
- & Quanshun Li
-
Article
| Open Access
Ligand-tethered lipid nanoparticles for targeted RNA delivery to treat liver fibrosis
Gene delivery to fibroblasts for liver fibrosis treatment remains challenging. Here the authors develop a combinatorial library of ligand-tethered lipidoids via a modular synthetic method and adopt a 2-round screening strategy to identify lipidoids for potent and selective gene delivery to fibroblasts.
- Xuexiang Han
- , Ningqiang Gong
- & Michael J. Mitchell
-
Article
| Open Access
Targeted systematic evolution of an RNA platform neutralizing DNMT1 function and controlling DNA methylation
Here the authors generate an RNA-based platform to neutralize the major epigenetic player DNMT1. Using this targeted approach, aberrant DNA methylation in cancer can be corrected.
- Carla L. Esposito
- , Ida Autiero
- & Annalisa Di Ruscio
-
Article
| Open Access
XNAzymes targeting the SARS-CoV-2 genome inhibit viral infection
RNA viruses have been responsible for large-scale epidemics and pandemics throughout the last few centuries. Here, the authors show the design, synthesis and screening of artificial RNA endonuclease XNAzymes capable of cleaving genomic SARS-CoV-2 RNA and self-assembling into enzymatic nanostructures inhibiting cellular viral replication.
- Pehuén Pereyra Gerber
- , Maria J. Donde
- & Alexander I. Taylor
-
Article
| Open Access
DNA-delivered antibody cocktail exhibits improved pharmacokinetics and confers prophylactic protection against SARS-CoV-2
Here, Parzych et al describe the development of a DNA-delivered SARS-CoV-2 mAb cocktail displaying synergistic RBD binding that confers broad neutralizing activity against variants of concern and prophylactic efficacy in multiple small animal models.
- Elizabeth M. Parzych
- , Jianqiu Du
- & David B. Weiner
-
Article
| Open Access
Chemical engineering of therapeutic siRNAs for allele-specific gene silencing in Huntington’s disease models
Chemically modified siRNAs distinguish between mutant and normal huntingtin based on a single nucleotide difference and lower mutant huntingtin specifically in patient derived cells and in a mouse model of Huntington’s disease.
- Faith Conroy
- , Rachael Miller
- & Edith L. Pfister
-
Article
| Open Access
Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo
Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands.
- Huanzhen Ni
- , Marine Z. C. Hatit
- & James E. Dahlman
-
Article
| Open Access
An LNA-amide modification that enhances the cell uptake and activity of phosphorothioate exon-skipping oligonucleotides
Oligonucleotides targeting mRNA are promising therapeutic agents but suffer from poor bioavailability. Here, the authors develop reduced-charge oligonucleotides with artificial LNA-amide linkages with improved cell uptake and minimal structural deviation to the DNA:RNA duplex.
- Ysobel R. Baker
- , Cameron Thorpe
- & Tom Brown
-
Article
| Open Access
Gene-specific nonsense-mediated mRNA decay targeting for cystic fibrosis therapy
The W1282X nonsense mutation in the CFTR gene causes cystic fibrosis by reducing its mRNA and functional protein levels. Here the authors developed antisense-oligonucleotide cocktails that restore CFTR protein function by gene-specific stabilization of CFTR mRNA.
- Young Jin Kim
- , Tomoki Nomakuchi
- & Adrian R. Krainer
-
Article
| Open Access
RNA aptamers specific for transmembrane p24 trafficking protein 6 and Clusterin for the targeted delivery of imaging reagents and RNA therapeutics to human β cells
Development of probes specific for human β-cells could aid in delivery of therapeutics and monitoring β-cells mass during diabetes progression or islet transplantation. Here the authors identify two RNA aptamers specific for β-cells that allow efficient transfection of human islets and β-cell quantification of human islet grafts in immunodeficient mice.
- Dimitri Van Simaeys
- , Adriana De La Fuente
- & Paolo Serafini
-
Article
| Open Access
Guide RNAs containing universal bases enable Cas9/Cas12a recognition of polymorphic sequences
Genetic variance poses a major challenge for CRISPR-based human therapeutics and pathogen diagnostics. Here the authors show how to circumvent this issue by using guide RNAs containing universal bases to tailor Cas9/Cas12 specificity.
- Amanda R. Krysler
- , Christopher R. Cromwell
- & Basil P. Hubbard
-
Article
| Open Access
MicroRNA-365 regulates human cardiac action potential duration
An abnormal cardiac action potential underlies different types of cardiac arrhythmias. Here the authors show that microRNA-365 regulates the cardiac action potential by modulating key cardiac repolarizing channels.
- Dena Esfandyari
- , Bio Maria Ghéo Idrissou
- & Stefan Engelhardt
-
Article
| Open Access
DNA/RNA heteroduplex oligonucleotide technology for regulating lymphocytes in vivo
Using gene silencing to regulate lymphocyte function is a promising therapeutic approach for autommunity, inflammation and cancer. Here the authors use a heteroduplex oligonucleotide for improved potency, efficacy and longer retention times.
- Masaki Ohyagi
- , Tetsuya Nagata
- & Takanori Yokota
-
Comment
| Open Access
An ionizable lipid toolbox for RNA delivery
RNA therapeutics have benefited significantly from decades of research on lipid nanoparticles, specifically its key component—the ionizable lipid. This comment discusses the major ionizable lipid types, and provides perspectives for future development.
- Xuexiang Han
- , Hanwen Zhang
- & Michael J. Mitchell
-
Article
| Open Access
GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain
GAPDH is generally considered a housekeeping gene and functions in glycolysis. Here, the authors show that GAPDH has a role in promoting vesicle clustering in endosomes and can load siRNA onto the surface of extracellular vesicles, which can be exploited therapeutically.
- Ghulam Hassan Dar
- , Cláudia C. Mendes
- & Matthew J. A. Wood
-
Article
| Open Access
A comparative analysis of cell surface targeting aptamers
Aptamers could potentially be widely used in therapy and diagnostics. Here the authors use standardised assay conditions to compare aptamer properties in tumour targeting.
- Linsley Kelly
- , Keith E. Maier
- & Matthew Levy
-
Article
| Open Access
Reprogrammed CRISPR-Cas13b suppresses SARS-CoV-2 replication and circumvents its mutational escape through mismatch tolerance
Cas13b can be harnessed to target and degrade RNA transcripts inside a cellular environment. Here the authors reprogram Cas13b to target SARSCoV-2 transcripts in infected mammalian cells and reveal its resilience to variants thanks to single mismatch tolerance.
- Mohamed Fareh
- , Wei Zhao
- & Joseph A. Trapani
-
Article
| Open Access
Nucleoside-modified VEGFC mRNA induces organ-specific lymphatic growth and reverses experimental lymphedema
Dysfunction of the lymphatic system leads to secondary lymphedema and results in degradation of quality of life. Here, the authors show that delivery of nucleoside-modified Vascular Endothelial Growth Factor C (VEGFC) mRNA, encapsulated in lipid nanoparticles, induces organ-specific lymphatic growth and reverses experimental lymphedema.
- Dániel Szőke
- , Gábor Kovács
- & Zoltán Jakus
-
Article
| Open Access
Programmably tiling rigidified DNA brick on gold nanoparticle as multi-functional shell for cancer-targeted delivery of siRNAs
Small interfering RNA (siRNA) is used to regulate gene expression for therapeutic purposes, but the design of stable and efficient siRNA delivery systems is challenging. Here, the authors develop a siRNA-encapsulated and aptamer-incorporated core/shell nanoparticle for controlled siRNA delivery, with high stability, tumor-specific targeting and long circulation time.
- Chang Xue
- , Shuyao Hu
- & Zai-Sheng Wu
-
Article
| Open Access
Variant-selective stereopure oligonucleotides protect against pathologies associated with C9orf72-repeat expansion in preclinical models
C9orf72 expansion mutations are the most common genetic cause of ALS and FTD, which have limited therapies. The authors generate stereopure oligonucleotides that selectively deplete expansion-containing transcripts and protect against expansion-associated pathologies in preclinical models.
- Yuanjing Liu
- , Jean-Cosme Dodart
- & Robert H. Brown Jr.
-
Article
| Open Access
Antisense oligonucleotide modulation of non-productive alternative splicing upregulates gene expression
Restoration of normal gene expression is one way to treat monogenic disorders. Here the authors target naturally occurring non-productive alternative splicing using antisense oligonucleotides to promote the production of functional proteins.
- Kian Huat Lim
- , Zhou Han
- & Isabel Aznarez
-
Article
| Open Access
Systemic nanoparticle delivery of CRISPR-Cas9 ribonucleoproteins for effective tissue specific genome editing
Therapeutic targets of CRISPR-Cas can often not be accessed due to lack of carriers to deliver RNPs systematically. Here, the authors engineer modified lipid nanoparticles for delivery of gene editing proteins to specific tissues.
- Tuo Wei
- , Qiang Cheng
- & Daniel J. Siegwart
-
Article
| Open Access
Development of an exon skipping therapy for X-linked Alport syndrome with truncating variants in COL4A5
Alport syndrome is a progressive inherited nephritis accompanied by sensorineural loss of hearing and ocular abnormalities, for which there is currently no effective therapy. Here, the authors develop an exon-skipping therapy using an antisense-oligonucleotide and show it is effective in mouse models.
- Tomohiko Yamamura
- , Tomoko Horinouchi
- & Kandai Nozu
-
Article
| Open Access
Imaging small molecule-induced endosomal escape of siRNA
Therapeutic siRNA becomes trapped in endosomes, limiting its efficacy. Here the authors use fluorescently-tagged galectin-9 as a biosensor for membrane damage to monitor endosomal escape of cholesterol-conjugated siRNA following treatment of small molecule membrane-destabilising drugs.
- Hampus Du Rietz
- , Hampus Hedlund
- & Anders Wittrup
-
Article
| Open Access
Inhibition of DNA damage response at telomeres improves the detrimental phenotypes of Hutchinson–Gilford Progeria Syndrome
Hutchinson–Gilford progeria syndrome causes premature aging. Here the authors show that activation of the DNA damage response at dysfunctional telomeres and transcription of telomeric non-coding RNAs contributes to the pathogenesis, which can be ameliorated by treatment with sequence-specific telomeric antisense oligonucleotides.
- Julio Aguado
- , Agustin Sola-Carvajal
- & Fabrizio d’Adda di Fagagna
-
Article
| Open Access
Intradermal delivery of modified mRNA encoding VEGF-A in patients with type 2 diabetes
Chemically modified mRNA is a new approach for therapeutic protein expression that could be applied to angiogenesis. Here the authors show in a phase 1 clinical trial that a modified mRNA encoding VEGF-A is well tolerated in patients with type 2 diabetes.
- Li-Ming Gan
- , Maria Lagerström-Fermér
- & Regina Fritsche-Danielson
-
Article
| Open Access
RNA inhibitors of nuclear proteins responsible for multiple organ dysfunction syndrome
Multiple organ dysfunction syndrome (MODS) is a serious event that can occur following infection or tissue injury, and is partly mediated by histones released in circulation. Here, the authors develop aptamers that neutralise histones involved in MODS, and demonstrate efficacy in human cells and in mouse models.
- Kevin T. Urak
- , Giselle N. Blanco
- & Paloma H. Giangrande
-
Article
| Open Access
Cell specific delivery of modified mRNA expressing therapeutic proteins to leukocytes
Therapeutic alteration of protein expression using modified mRNA is limited by immunogenicity and instability in vivo. Here the authors use antibody-coated lipid nanoparticles to deliver mRNA to leukocytes and drive expression of anti-inflammatory cytokines in an inflammatory bowel disease mouse model.
- Nuphar Veiga
- , Meir Goldsmith
- & Dan Peer
-
Article
| Open Access
A RNA producing DNA hydrogel as a platform for a high performance RNA interference system
Interfering RNA have a range of therapeutic and research based applications, issues with delivery have made systems that make siRNA in situ of interest. Here, the author report on the creation of a DNA hydrogel with improved stability and transcription efficiency over plasmid DNA.
- Jaejung Song
- , Minhyuk Lee
- & Nokyoung Park
-
Article
| Open Access
Engineering circular RNA for potent and stable translation in eukaryotic cells
Circular RNAs have recently been shown to have protein-coding potential. Here the authors design a self-splicing RNA that, when circularized, provides for stable high-yield protein production.
- R. Alexander Wesselhoeft
- , Piotr S. Kowalski
- & Daniel G. Anderson
-
Article
| Open Access
Heavily and fully modified RNAs guide efficient SpyCas9-mediated genome editing
Resistance of gRNA to ubiquitous ribonucleases is required for CRISPR-Cas9-based therapeutics. Here, the authors explore chemical modifications at all positions of the crRNA guide and tracrRNA cofactor, and identify modified versions that are more potent and stable than their unmodified counterparts in editing human cells.
- Aamir Mir
- , Julia F. Alterman
- & Erik J. Sontheimer
-
Article
| Open Access
In utero nanoparticle delivery for site-specific genome editing
The correction of genetic defects in utero could allow for improved outcomes of gene therapy. Here, the authors demonstrate safe delivery of nanoparticles to fetal mouse tissues, and show that nanoparticles containing peptide nucleic acids to edit the beta-globin gene are effective in a mouse model of beta-thalassemia.
- Adele S. Ricciardi
- , Raman Bahal
- & W. Mark Saltzman
-
Article
| Open Access
HSP27 is a partner of JAK2-STAT5 and a potential therapeutic target in myelofibrosis
Myelofibrosis is a chronic degenerative disorder characterized by progressive bone marrow fibrosis. Here, the authors show that the chaperone HSP27 contributes to myelofibrosis via regulation of the JAK2/STAT5 pathway, and that antisense oligonucleotides targeting HSP27 are effective in two mouse models of the disease
- Margaux Sevin
- , Lucia Kubovcakova
- & Aurélie de Thonel
-
Article
| Open Access
Selection of GalNAc-conjugated siRNAs with limited off-target-driven rat hepatotoxicity
A subset of chemically-modified siRNAs conjugated to trivalent GalNAc may fail during nonclinical development due to rat hepatotoxicity. Here, the authors show that hepatotoxicity may be accounted for by microRNA-like off-target effects of siRNA and can be mitigated by a thermally destabilizing modification in the siRNA seed region.
- Maja M. Janas
- , Mark K. Schlegel
- & Vasant Jadhav
-
Article
| Open Access
Regulation of chitinase-3-like-1 in T cell elicits Th1 and cytotoxic responses to inhibit lung metastasis
Chitinase-3-like-1 (Chi3l1) has been involved in inflammation and pulmonary metastasis. Here the authors show that Chi3l1 inhibits the T cell response by negatively regulating their activation and that, in a mouse model of melanoma, T cell-targeted silencing of Chi3l1 results in reduced lung metastasis.
- Do-Hyun Kim
- , Hong-Jai Park
- & Je-Min Choi
-
Article
| Open Access
Amphiphilic nanocarrier-induced modulation of PLK1 and miR-34a leads to improved therapeutic response in pancreatic cancer
Treatment of pancreatic ductal adenocarcinoma is still challenging and patients survival has only marginally improved in the last decade. Here the authors produce a PGA-based polymeric nanocarrier for the dual delivery of miR-34a-mimic and PLK1-targeting siRNA resulting in killing of pancreatic cancer cells in vivo.
- Hadas Gibori
- , Shay Eliyahu
- & Ronit Satchi-Fainaro
-
Article
| Open Access
Antisense oligonucleotide-mediated Dnm2 knockdown prevents and reverts myotubular myopathy in mice
X-linked myotubular myopathy is caused by mutations in the gene coding for myotubularin 1, and is characterized by overexpression of dynamin 2. Here the authors develop antisense oligonucleotides to dynamin 2, and show that systemic injection leads to improved pathology in mice.
- Hichem Tasfaout
- , Suzie Buono
- & Jocelyn Laporte
-
Article
| Open Access
Light-inducible antimiR-92a as a therapeutic strategy to promote skin repair in healing-impaired diabetic mice
Inhibition of microRNAs using antimiRs is a potential therapeutic option for a number of diseases, but systemic inhibition may cause adverse effects. Here the authors develop light-activated antimiRs directed against miR-92a, and show localized inhibition in the skin and improved wound healing in diabetic mice.
- Tina Lucas
- , Florian Schäfer
- & Stefanie Dimmeler
-
Article
| Open Access
Administration of nucleoside-modified mRNA encoding broadly neutralizing antibody protects humanized mice from HIV-1 challenge
Monoclonal antibodies are highly effective therapeutics that can be delivered as proteins or encoded DNA or mRNA. Here the authors develop lipid nanoparticle-formulated nucleoside-modified mRNA encoding an HIV-1 neutralizing antibody and see sustained and protective antibody levels in treated mice.
- Norbert Pardi
- , Anthony J. Secreto
- & Drew Weissman
-
Article
| Open Access
Small RNA zippers lock miRNA molecules and block miRNA function in mammalian cells
MicroRNAs regulate a wide range of biological processes and being able to inhibit their function could allow the development of therapeutic options. Here the authors describe a ‘small RNA zipper’ that sequesters miRNAs by forming a chain of DNA:RNA duplexes.
- Lingyu Meng
- , Cuicui Liu
- & Zuoren Yu
Inhibition of iRhom1 by CD44-targeting nanocarrier for improved cancer immunochemotherapy