RNA Interference by Single- and Double-stranded siRNA With a DNA Extension Containing a 3′ Nuclease-resistant Mini-hairpin Structure

  title={RNA Interference by Single- and Double-stranded siRNA With a DNA Extension Containing a 3′ Nuclease-resistant Mini-hairpin Structure},
  author={S. Allison and J. Milner},
  journal={Molecular Therapy. Nucleic Acids},
Selective gene silencing by RNA interference (RNAi) involves double-stranded small interfering RNA (ds siRNA) composed of single-stranded (ss) guide and passenger RNAs. siRNA is recognized and processed by Ago2 and C3PO, endonucleases of the RNA-induced silencing complex (RISC). RISC cleaves passenger RNA, exposing the guide RNA for base-pairing with its homologous mRNA target. Remarkably, the 3′ end of passenger RNA can accommodate a DNA extension of 19-nucleotides without loss of RNAi… Expand
DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization
This work uses DNA oligomers as adaptors to increase size and charge of cargo siRNA, resulting in increased polyplex stability, which in turn boosts transfection efficiency and shows that DNA adaptors provide the opportunity to readily link additional functional domains to siRNA. Expand
Anti-c-myc RNAi-Based Onconanotherapeutics
The various anti-c-myc RNAi-based nanosystems that have come to the fore, especially between 2005 and 2020, are reviewed. Expand
Rekindling RNAi Therapy: Materials Design Requirements for In Vivo siRNA Delivery.
The main aims is to identify and describe key design features needed for nanoscale vehicles to achieve effective delivery of siRNA-mediated gene silencing agents in vivo. Expand
Helper lipid structure influences protein adsorption and delivery of lipid nanoparticles to spleen and liver.
Understanding the role of the helper lipid in LNP biodistribution and ApoE adsorption may aid in the future design of LNPs for nucleic acid therapeutics. Expand
Selective in vitro anti-cancer activity of non-alkylating minor groove binders.
An initial investigation into the mechanism of action of one compound from this class of Strathclyde Minor Groove Binders shows that neither DNA double strand breaks nor the DNA damage stress sensor protein p53 are induced, indicating that these S-MGBs are unlikely to act through an alkylating or DNA damage response mechanism. Expand
Transfection types, methods and strategies: a technical review
With the vast choices of approaches available, it is hoped that this review will help researchers, especially those new to the field, in their decision making over the transfection protocol or strategy appropriate for their experimental aims. Expand
From influenza to COVID-19: Lipid nanoparticle mRNA vaccines at the frontiers of infectious diseases
This manuscript timely reviews the most advanced mRNA-LNP vaccines that have just been approved for emergency use and are in clinical trials, with a focus on the remarkable development of several COVID-19 vaccines, faster than any other vaccine in history. Expand
Delivery technologies to engineer natural killer cells for cancer immunotherapy.
An overview of the use of NK cells in cancer immunotherapy, their current status in clinical trials, as well as the design and implementation of delivery technologies-including viral, non-viral, and nanoparticle-based approaches-for engineering NK cell-based immunotherapies are provided. Expand
The Warburg effect as a therapeutic target for bladder cancers and intratumoral heterogeneity in associated molecular targets
Analysis of bladder cancer cell metabolic reprogramming to rely on aerobic glycolysis (the Warburg effect) and expression of associated molecular therapeutic targets by bladder cancer cells of different stages and grades, and in freshly resected clinical tissue, indicates that the Warberg effect is a feature of both non-muscle- and muscle- invasive bladder cancers. Expand
Stereochemical and pharmacological properties of phosphorothioate siRNAs


A bi-functional siRNA construct induces RNA interference and also primes PCR amplification for its own quantification
It is shown that the modified siRNA-DNA construct (termed ‘crook’ siRNA) functions as a primer for the PCR and described a novel, yet simple PCR protocol for its quantification (amolar levels/cell). Expand
Efficient Reduction of Target RNAs by Small Interfering RNA and RNase H-dependent Antisense Agents
Results demonstrate that siRNA oligonucleotide- and RNase H-dependent antisense strategies are both valid strategies for evaluating function of genes in cell-based assays. Expand
mRNA knockdown by single strand RNA is improved by chemical modifications
It is identified that 2′F ribose modifications coupled with 5′-end phosphorylation vastly improves ssRNA activity both in vitro and in vivo, suggesting ssRNA may operate through a mechanism beyond conventional Ago2 slicer activity. Expand
Conversion of pre-RISC to holo-RISC by Ago2 during assembly of RNAi complexes.
It is demonstrated in vivo that efficient removal of the passenger strand from RISC requires the cleavage activity of Ago2, and a new intermediate complex is identified in the RISC assembly pathway, pre-RISC, in which Ago2 is stably bound to double-stranded siRNA. Expand
Nuclease resistance of an extraordinarily thermostable mini-hairpin DNA fragment, d(GCGAAGC) and its application to in vitro protein synthesis.
By using this method, dihydrofolate reductase (DHFR) mRNA was stabilized against nucleases contaminating a cell-free translation system of E.coli, with a consequent increase in protein synthesis efficiency of 200%. Expand
C3PO, an Endoribonuclease That Promotes RNAi by Facilitating RISC Activation
An in vitro RNAi reconstitution system is established and C3PO is identified as a key activator of the core RNAi machinery, a complex of Translin and Trax. Expand
Structure of an argonaute silencing complex with a seed-containing guide DNA and target RNA duplex
Consistent with the geometry of the ternary complex, bulges residing in the seed segments of the target, but not the guide strand, were better accommodated and their complexes were catalytically active. Expand
Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes
The slicer activity of the RNA-induced silencing complex resides within its Argonaute (Ago) component, in which the PIWI domain provides the catalytic residues governing guide-strand mediatedExpand
On the road to reading the RNA-interference code
The finding that sequence-specific gene silencing occurs in response to the presence of double-stranded RNAs has had an enormous impact on biology, uncovering an unsuspected level of regulation ofExpand
Structure of the guide-strand-containing argonaute silencing complex
The crystal structure of Thermus thermophilus argonaute bound to a 5′-phosphorylated 21-base DNA guide strand is reported on, thereby identifying the nucleic-acid-binding channel positioned between the PAZ- and PIWI-containing lobes, as well as the pivot-like conformational changes associated with complex formation. Expand