article{5993109, author = {De Backer, Lynn and Braeckmans, Kevin and Stuart, Marc CA and Demeester, Jo and De Smedt, Stefaan and Raemdonck, Koen}, issn = {0168-3659}, journal = {JOURNAL OF CONTROLLED RELEASE}, keyword = {DRUG-DELIVERY,EXOGENOUS SURFACTANT,LUNG,THERAPEUTICS,THERAPIES,SMALL-INTERFERING RNA,LOADED DEXTRAN NANOGELS,POLYMER HYBRID NANOPARTICLES,Targeted delivery,Pulmonary surfactant,siRNA delivery,Lung,Hybrid nanoparticles,Dextran nanogels,PLATFORM,CARRIERS}, language = {eng}, pages = {177--186}, title = {Bio-inspired pulmonary surfactant-modified nanogels: a promising siRNA delivery system}, url = {http://dx.doi.org/10.1016/j.jconrel.2015.03.015}, volume = {206}, year = {2015 ...
RNA interference (RNAi) is a gene-silencing mechanism by which a ribonucleoprotein complex, the RNA-induced silencing complex (RISC) and a double-stranded (ds) short-interfering RNA (siRNA), targets a complementary mRNA for site-specific cleavage and subsequent degradation. While longer dsRNA are endogenously processed into 21- to 24-nucleotide (nt) siRNAs or miRNAs to induce gene silencing, RNAi studies in human cells typically use synthetic 19- to 20-nt siRNA duplexes with 2-nt overhangs at the 3-end of both strands. Here, we report that systematic synthesis and analysis of siRNAs with deletions at the passenger and/or guide strand revealed a short RNAi trigger, 16-nt siRNA, which induces potent RNAi in human cells. Our results indicate that the minimal requirement for dsRNA to trigger RNAi is an approximately 42 A A-form helix with approximately 1.5 helical turns. The 16-nt siRNA more effectively knocked down mRNA and protein levels than 19-nt siRNA when targeting the endogenous CDK9 gene,
DESCRIPTION (provided by applicant): The proposed research will enable mammalian cells to be used in mutation/selection experiments to identify genes involved in cellular processes. The objective of the proposal is to develop procedures for preparing siRNA libraries using genomic DNA and cDNA samples as the source material for siRNA templates. Libraries of siRNA vectors could comprise 1,000,000,000 to 1,000,000,000,000 unique siRNA expression vectors, making it possible to cover entire mammalian genomes with overlapping siRNAs. Mammalian cell populations transfected or transduced with siRNA vector libraries will be placed under selection to create cell populations expressing a desirable phenotype. The siRNA vector(s) integrated in the genomes of the selected cells will be amplified, cloned, and sequenced to reveal the siRNA template sequence. The siRNA sequences will be used to identify the genes whose down-regulation create the phenotype that was selected. The siRNA vector libraries should ...
The success of siRNA-based therapeutics highly depends on a safe and efficient delivery of siRNA into the cytosol. In this study, we post-modified the primary amines on dendritic polyglycerolamine (dPG-NH2) with different ratios of two relevant amino acids, namely, arginine (Arg) and histidine (His). To investigate the effects from introducing Arg and His to dPG, the resulting polyplexes of amino acid functionalized dPG-NH2s (AAdPGs)/siRNA were evaluated regarding cytotoxicity, transfection efficiency, and cellular uptake. Among AAdPGs, an optimal vector with (1:3) Arg to His ratio, showed efficient siRNA transfection with minimal cytotoxicity (cell viability ≥ 90%) in NIH 3T3 cells line. We also demonstrated that the cytotoxicity of dPG-NH2 decreased as a result of amino acid functionalization. While the incorporation of both cationic (Arg) and pH-responsive residues (His) are important for safe and efficient siRNA transfection, this study indicates that AAdPGs containing higher degrees of ...
TY - CHAP. T1 - Short hairpin RNA-mediated gene silencing. AU - Lambeth, Luke S. AU - Smith, Craig A.. PY - 2013. Y1 - 2013. N2 - Since thefirst application of RNA interference (RNAi) in mammalian cells, the expression of short hairpin RNAs (shRNAs) for targeted gene silencing has become a benchmark technology. Using plasmid and viral vectoring systems, the transcription of shRNA precursors that are effectively processed by the RNAi pathway can lead to potent gene knockdown. The past decade has seen continual advancement and improvement to the various strategies that can be used for shRNA delivery, and the use of shRNAs for clinical applications is well underway. Driving these developments has been the many benefits afforded by shRNA technologies, including the stable integration of expression constructs for long-term expression, infection of difficultto-target cell lines and tissues using viral vectors, and the temporal control of shRNA transcription by inducible promoters. The use of different ...
The time course of lipophilic siRNA accumulation in KB-8-5 cells. The incubation time after carrier-free transfection of cholesterol-conjugated siMDR with hexyl
As the portfolio of RNAi methods continues to expand, options become available for even the most complex systems being studied. Until recently, synthetic siRNA was the RNAi vehicle most broadly applicable to a wide variety of systems and applications. With commercial suppliers designing and producing synthetic siRNAs, little manipulation is required for the consumer. This format is amenable to any scale of research being performed provided the system is easily transfected (e.g., standard transformed cell lines). However, obstacles for using synthetic siRNAs include being a non-renewable resource, the transient nature of silencing, and the difficulty faced in transfecting primary cells and non-dividing cell lines such as neurons, lymphocytes, and macrophages. In addition, in vivo knockdown studies are particularly cumbersome.. For those facing the above hurdles, DNA vector-based shRNA methods provide the necessary solutions. shRNA expression vectors may be propagated in Escherichia coli and, ...
Results Green fluorescence was observed in the cells transfected of negative control siRNA group though the fluorescence microscope. Compared with the blank control group, The MTT assay determined that the survival rate of H9C2 was decreased (p , 0.05) after the injured by hypoxia. And the results of flow cytometry showed that hypoxia increased cell apoptosis rate (P , 0.01) and the concentration of calcium (p , 0.01), while the transfection of Bim-siRNA reduced the effects caused by hypoxia (P , 0.05 or P , 0.01). Compared with the hypoxia group, the transfection of Bim-siRNA increased the cell survival rate, decreased cell apoptosis rate and the concentration of calcium (p , 0.01 or p , 0.05). While there was no significant difference among Hypoxia Group, Hypoxia + Negative Control siRNA Group and Hypoxia + Mock control Group (p , 0.05). The results of Western blotting showed that the transfection of Bim-siRNA reduced the expression of Bim obviously (p , 0.01); meanwhile, reduced the ...
The most enjoyable part in following RNAi Therapeutics is to look at the rich stream of scientific data and determine the absolute maturity and competitive position of the technologies and companies involved, as well as getting a glimpse at relationship dynamics. I therefore thought to share today two examples of this that I picked up recently. One is a paper by Sirna Therapeutics/Merck shedding some light on their approach towards RNAi pharmacology and RNAi trigger design. The other is some intriguing evidence that Silence Therapeutics most important gene target, PKN3, is gaining traction in the pharmaceutical space. Studying the pharmacology of siRNA delivery. Pei and colleagues from Merck published in RNA a nice paper on better understanding the pharmacology of siRNA delivery [Pei et al. (2010). Quantitative evaluation of siRNA delivery in vivo]. Unlike small molecules or even antibodies, the pharmacology of RNAi Therapeutics is more complex as simply measuring the raw tissue abundance of an ...
Semple, SC, Akinc, A, Sandhu, A, Mui, B, Chow, C, Sah, D, Stebbing, D, Crosley, E, Hafez, I, Dorkin, JR, Qin, J, Lam, K, Wong, K, Nechev, L, Eisenhardt, ML, Jayaraman, M, Kazem, M, Maier, M, Srinivasulu, M, Weinstein, M, Chen, Q, Alvarez, R, Barros, S, Klimuk, SK, Borland, T, Kosovrasti, V, Tam, Y, MacLachlan, I, Manoharan, M, Ciufolini, MA, Tracy, M, de Fougerolles, A, Cullis, PR, Madden, TD, Hope, ...
mTOR and Rictor mediate cell migration.A: siRNA-induced mTOR depletion inhibits Tsc2−/− MEF migration. Tsc2−/− MEFs were transfected with siRNA mTOR or
Researchers working in the field of siRNA based therapeutics for viruses have generated a vast amount of data over the years. However, bioinformatics resources in the field were lacking and there was no viral siRNA efficacy prediction method available. In this direction, we have recently developed a comprehensive viral siRNA database "VIRsiRNAdb" [48] and another "HIVsirDB" [49] exclusively for HIV. Now we have developed VIRsiRNApred -a viral siRNA efficacy prediction algorithm.. Although many mammalian siRNA prediction algorithms have been developed in the past [33], these methods either classify a siRNA as effective/non-effective [29] or predict the inhibition efficacy of a siRNA [31, 32]. However, there is limited success in predicting siRNA efficacy due to limited size and diversity of available siRNA datasets [50].. Mammalian siRNA efficacy prediction methods were initially developed using siRNA tested under heterogeneous experimental conditions like Saetrom (581 siRNA), Shabalina (653 ...
GAITHERSBURG, Md., June 29 /PRNewswire/ -- Sirnaomics Receives Two SBIR Grants from NIH for Developing siRNA Therapeutics to Treat Glioblastoma and...
Suzhou Ribo Life Sciences is developing siRNA therapeutics for a range of diseases of high unmet need in China, including liver fibrosis, HIV, liver and
You can add the following controls to your FlexiPlate siRNA plate: AllStars Negative Control siRNA, AllStars Cell Death Control siRNA, Negative Control siRNA, Human GAPDH siRNA, Human Beta-Actin siRNA, Human and mouse MAPK1 siRNA, Human or mouse Lamin A/C siRNA, Mouse AKT1 siRNA, or other siRNAs from GeneGlobe, such as HP Validated siRNAs. ...
RNA interference pathways can involve amplification of secondary siRNAs by RNA-dependent RNA polymerases. In plants, RDR6-dependent secondary siRNAs arise from transcripts targeted by some microRNAs (miRNAs). Here, Arabidopsis thaliana secondary siRNAs from mRNA as well as trans-acting siRNAs are sh …
One siRNA sequence, many cell lines - posted in siRNA, microRNA and RNAi: Hi all, Im new to process of siRNA transfection and I was wondering: Will one siRNA sequence (previously validated in the lab) be good enough to transfect multiple other cell lines from the same organism? I understand that the actual process of transfection will be different for each cell line, but I am curious as to whether I need to also worry about the sequence itself. Thanks!
Our study supports a previous conjecture that primary RdDM is required to initiate 24‐nt secondary siRNA formation. This requirement was initially suggested by the absence of secondary siRNAs in nrpe1 and drd1 mutants, which still produce primary siRNAs and the overlapping nascent RNA but lack primary RdDM (Kanno et al, 2008). The nascent RNA stably accumulates in the absence of primary RdDM in non‐silenced plants but, in that instance, is presumably a Pol II transcript (Figure 6) because it is still made in an nrpd1 mutant. The key function of primary RdDM appears to be in attracting the secondary siRNA‐generating machinery, which includes Pol IV and RDR2. The contribution may be direct if primary RdDM provides a template that is preferentially recognized by Pol IV, which has been proposed to transcribe methylated DNA (Herr et al, 2005; Onodera et al, 2005). In this model, Pol IV would transcribe the nascent RNA from the methylated template (Figure 6). An indirect contribution of primary ...
The field of RNA-based gene regulation has been attracting increasing interest over the past couple of years, and the regulation of gene expression by small dsRNAs is being studied intensively. Such interference can be mediated by siRNAs, which cleave a sequence-specific target mRNA, or by micro-RNAs, which inhibit translation of a target mRNA. Noncoding RNAs have also been found to play important roles in the regulation of gene expression, for example, in gene silencing by methylation of DNA or histones. Small interfering RNAs are expected to have medical application in human therapy as drugs with high specificity for their molecular targets.. A number of studies on synthetic siRNAs or DNA vector-derived small hairpin RNAs (shRNAs) in cell culture systems have been published, and there are also several animal studies (15, 16, 17, 18, 19) . McCaffrey et al. (15) cotransfected the firefly luciferase gene along with synthetic siRNAs or a shRNA expression vector into mice by hydrodynamic injection ...
In recent years, small interference RNAs (siRNAs) have greatly enhanced our understanding of protein functions by allowing knockdown of targeted proteins at the mRNA level
MISSION® siRNA Universal Negative Controls are an essential component to any siRNA experiment. Using a Negative siRNA control allows the researcher to create a baseline for mRNA knockdown efficiency. MISSION siRNA Universal Negative Controls have been tested in human, rat, and mouse cells. All have carefully designed to have no homology to known gene sequences.

Benefits of MISSION Universal Negative siRNA Control:
  • Universal controls designed for use in a wide variety of species including Human, Mouse, Rat, Bovine, Pig, Chicken, Sheep, and CHO cells.
  • Novel design method used to ensure no homology to all Mature and Predicted RefSeq mRNA sequences.
  • Contain no known immune-response motifs.1
  • The unconjugated controls were validated with Agilent 40K human gene arrays to ensure no significant nonspecific gene interactions.
  • Functionally validated to show no immune response via qRT PCR
  • Two different Universal Negative Control
Due to reasons discussed in the previous post, almost all of these programs are Direct RNAi programs, i.e. the siRNA is administered close to the diseased site. All except for Acuitys program are also with siRNAs that have been chemically modified (to enhance stability etc), and it remains to be seen whether Acuitys strategy to plunge into the clinic first was a wise one. Sirna Therapeutics soon followed suit, but I think took the right decision to invest time to carefully think about how to position their potential product in the more and more crowded AMD market. Sirna Therapeutics was also the first public company based on RNAi. Formerly known as Ribozyme Pharmaceuticals, they leveraged their experience with RNAs to build an operation that had all the tools to to build a decent IP portfolio and quickly enter the clinic. This IP portfolio is mostly based on chemistry and targeting many genes one by one such that it could claim exclusivity for targeting those genes with RNAi. It remains to be ...
Benenson and colleagues engineered a target gene to be sensitive to several different siRNAs of their own design. In the simplest case, they introduced a single siRNA molecule to switch off a target gene that encoded a fluorescent protein. In more complex cases, a pair of siRNAs or either of two siRNAs switched off another target gene, which in turn switched off a gene for a fluorescent protein. To make sure the system worked as intended, the researchers based their siRNAs on those of other species, they report in a paper published online today by Nature Biotechnology ...
Download Free Full-Text of an article RORC2 GENE SILENCING IN HUMAN TH17 CELLS BY SIRNA: DESIGN AND EVALUATION OF HIGHLY EFFICIENT SIRNA
Anew detection mechanism for the control of successful siRNA delivery to cells or tissue is introduced using a siRNA-based probe that is capable of inducing a ...
PlanningOperationsProduct ManagementProductionPublic RelationsResearchSalesOther Yes, I need to discuss cells from Adweek about programs, formats and entries that they are may regulate of siRNA Design: Methods and Protocols to me. You emit together randomized to this siRNA Design: Methods. puncture us to find up to siRNA realize as completed to this selection.
siRNA transfection is a powerful tool used to understand the mechanisms of gene regulation and molecular pathways. The following 10 tips will help you to optimize your siRNA transfection.
RNA interference (RNAi) is a powerful tool in the study of gene function. We added poly(A) tails to the 3 ends of siRNA antisense strands by in vitro..
Acquired by Silicon Image Anchor Bay Technologies is developing a new class of components and systems, based on high-performance Digital Video and Audio format-conversion technologies, for the Digital A/V market.. ...
Transcription factor; can act both as activator and as repressor. Binds the 5-CACCC-3 core sequence. Binds to the promoter region of its own gene and can activate its own transcription. Regulates the expression of key transcription factors during embryonic development. Plays an important role in maintaining embryonic stem cells, and in preventing their differentiation. Required for establishing the barrier function of the skin and for postnatal maturation and maintenance of the ocular surface. Involved in the differentiation of epithelial cells and may also function in skeletal and kidney development. Contributes to the down-regulation of p53/TP53 transcription. (Source: UniProt http://www.uniprot.org/uniprot/o43474 ...
Techniques that regulate the level of protein expression by targeting genes at the DNA or RNA level have proven to be powerful strategies in the drive to understand protein expression and function. However, because of their very nature, these techniques are restricted in terms of speed, specificity and reversibility. For instance, these genetic methods of disrupting protein expression can take days to weeks; consequently, cellular and molecular compensation may occur, thereby obscuring expected phenotypes. In addition, as these genetic manipulations result in the eradication of all mRNA splice isoforms, as well as post-translationally modified versions of targeted proteins, these methods lack specificity and are largely limited to studying context-dependent protein function. Finally, the reversibility of these genetic manipulations, including many recently developed on-and-off inducible methods, is relatively slow (being achieved on a timescale of days to weeks) and is incomplete. These ...
SiRNAs exert their biological effect by guiding the degradation of their cognate mRNA sequence, thereby shutting down the corresponding protein production (gene silencing by RNA interference or RNAi). Due to this property, siRNAs are emerging as promising therapeutic agents for the treatment of inherited and acquired diseases, as well as research tools for the elucidation of gene function in both health and disease. Because of their lethality and prevalence, lung diseases have attracted particular attention as targets of siRNA-mediated cures. In addition, lung is accessible to therapeutic agents via multiple routes, e.g., through the nose and the mouth, thus obviating the need for targeting and making it an appealing target for RNAi-based therapeutic strategies. The clinical success of siRNA-mediated interventions critically depends upon the safety and efficacy of the delivery methods and agents. Delivery of siRNAs relevant to lung diseases has been attempted through multiple routes and using various
2345 Small interfering RNA (siRNA) oligonucleotides have been shown to be potent mediators of RNA interference (RNAi) that induce gene silencing with a high degree of sequence specificity. This has resulted in a rapid shift from antisense and ribozymes to siRNA for gene knockdown studies, and thus sparked strong interest in their use as therapeutics. Therapeutic use of siRNA, however, requires both improved biological stability and intracellular targeting. Thus far, gene inhibition by synthetic siRNA following intravenous administration has been limited to high pressure administration yielding primary effects in liver or using lipoplexes yielding primary effects in lung, in both cases via poorly understood mechanisms not readily adaptable to any other tissue and lacking clinical applicability. Recent work with aqueous siRNA administered parenterally by many routes found it could elicit effects on implanted tumors but without correlation to tumor exposure or tumor gene inhibition, leading the ...
Small interfering RNAs (siRNAs) are now established as the preferred tool to inhibit gene function in mammalian cells yet trigger unintended gene silencing due to their inherent miRNA-like behavior. Such off-target effects are primarily mediated by the sequence-specific interaction between the siRNA seed regions (position 2-8 of either siRNA strand counting from the 5-end) and complementary sequences in the 3UTR of (off-) targets. It was previously shown that chemical modification of siRNAs can reduce off-targeting but only very few modifications have been tested leaving more to be identified. Here we developed a luciferase reporter-based assay suitable to monitor siRNA off-targeting in a high throughput manner using stable cell lines. We investigated the impact of chemically modifying single nucleotide positions within the siRNA seed on siRNA function and off-targeting using 10 different types of chemical modifications, three different target sequences and three siRNA concentrations. We found ...
Four pre-designed shRNA constructs targeting cytoskeleton associated protein 2 (CKAP2), transcript variant 1 and one scrambled control. Each shRNA construct is driven by the U6 promoter and contains a GFP reporter.
Four pre-designed shRNA constructs targeting meiosis-specific nuclear structural 1 (MNS1) and one scrambled control. Each shRNA construct is driven by the U6 promoter and contains a GFP reporter.
An arrayed siRNA collection targeting mouse transcription factors. siGENOME siRNA is a cost-effective choice for RNAi screening. Available as SMARTpool or 4 individual siRNA reagents.
An arrayed siRNA collection targeting mouse genes in apoptosis pathways. siGENOME siRNA is a cost-effective choice for RNAi screening. Available as SMARTpool or 4 individual siRNA reagents.
Biology is generating more data than ever before. had been extracted from database magazines in BMC BMC and Bioinformatics Biology. To date, there were 19 significant contributors towards the task, each of whom continues to be shown Heparin sodium IC50 as an writer upon this publication. This task was taken up to highlight the grouped community facet of the MB project. The homepage continues to be visited 100 approximately?000 times. The task has 80 new users in total, and there were 15 approximately?000 edits. We wish that with ongoing improvements and through elevated publicity, use shall continue steadily to grow. Continue Reading. ...
Interleukin 17A (IL-17A) continues to be connected with protective instead of pathogenic response in Chagas disease (ChD). 42.1% of these were man. The Credit card group included 145 sufferers, which 58.6% were man, with ages which range from 23 to 67 years (mean of 49). The IND group shown higher degrees of IL-17A significantly, median of 26.16 (3.66C48.33) when compared with both the Credit card group, median of 13.89 (3.87C34.54) (. Continue Reading. ...
British biotech MiNA Therapeutics innovative RNA treatment may enhance liver cancer patients response to standard cancer therapies, suggest early results from a Phase I/IIa clinical trial.
The participants had Finnish as their native language and were from families with two parents and one to three children. For 18 of the families, at least one parent had either a bachelors (or equivalent), masters, or doctoral degree and for the majority of the families their monthly income was at or above the Finnish average level. The parents were asked about their childs possible hearing difficulties and other illnesses. The parents also provided the childs health summary, which contained information from the childs regular visits to a nurse and/or medical doctor that had occurred at least three times per year. Except for allergies, atopic skin or asthma, the subjects had no illnesses and no reported hearing or other. medical problems. The children were born at full term, had. normal birth weights, and their weight and height had developed normally. All of the children also had some Etoposide nmr musical experience outside the home as they had all attended the same playschool involving ...
An siRNA transfection reagent for efficient gene silencing in living cells via membrane fusion | Suitable for primary cells and non-dividing cells
/PRNewswire/ -- The RNAi Therapeutics Market (2nd Edition), 2019 - 2030 report has been added to ResearchAndMarkets.coms offering. This report features an...
Prime Standard QIA a leading supplier of products and technologies for nucleic acid separation, purification and handling today announced that it has launched a series of off-the-shelf human library siRNA sets, covering a broad gene family selection including kinases, GPCRs, apoptosis related genes, and oncogenes, to name a few.
IMO I think that people go with siRNA first to see if their siRNA sequence works and you get good knockdown. If you get good knowndown, then you can make the shRNA plasmid. But if you dont see good knockdown you end up making a new siRNA. If you went to shRNA first and then found that there is no knowndown, then it is a waste of time and money.. ...
ITGA5 - ITGA5 - Human, 4 unique 29mer shRNA constructs in retroviral GFP vector shRNA available for purchase from OriGene - Your Gene Company.
PDCD7 - Pdcd7 - Mouse, 4 unique 29mer shRNA constructs in retroviral GFP vector shRNA available for purchase from OriGene - Your Gene Company.
RNAi is a widely used methodology for gene silencing. The action mechanism of siRNA molecules has been well studied in recent years, and the technique..
This is the second post in a series, sharing a number of observed anti-patterns and corresponding patterns on the topic of organisational agility (aka digital transformation). We are in the midst of…