scholarly journals High-Pressure-Sprayed Double Stranded RNA Does Not Induce RNA Interference of a Reporter Gene

2020 ◽  
Vol 11 ◽  
Author(s):  
Veli Vural Uslu ◽  
Alexandra Bassler ◽  
Gabi Krczal ◽  
Michael Wassenegger
Keyword(s):  
High Pressure ◽  
Rna Interference ◽  
Reporter Gene ◽  
Plant Cell Wall ◽  
Defense Mechanism ◽  
Green Fluorescence Protein ◽  
Micro Rna ◽  
Small Rna Sequencing ◽  
Rna Molecules ◽  
Interfering Rna

In plants, RNA interference (RNAi) is an effective defense mechanism against pathogens and pests. RNAi mainly involves the micro RNA and the small interfering RNA (siRNA) pathways. The latter pathway is generally based on the processing of long double stranded RNAs (dsRNA) into siRNAs by DICER-LIKE endonucleases (DCLs). SiRNAs are loaded onto ARGONAUTE proteins to constitute the RNA-induced silencing complex (RISC). Natural dsRNAs derive from transcription of inverted repeats or of specific RNA molecules that are transcribed by RNA-directed RNA polymerase 6 (RDR6). Moreover, replication of infecting viruses/viroids results in the production of dsRNA intermediates that can serve as substrates for DCLs. The high effectiveness of RNAi both locally and systemically implicated that plants could become resistant to pathogens, including viruses, through artificial activation of RNAi by topical exogenous application of dsRNA. The most preferable procedure to exploit RNAi would be to simply spray naked dsRNAs onto mature plants that are specific for the attacking pathogens serving as a substitute for pesticides applications. However, the plant cell wall is a difficult barrier to overcome and only few reports claim that topical application of naked dsRNA triggers RNAi in plants. Using a transgenic Nicotiana benthamiana line, we found that high-pressure-sprayed naked dsRNA did not induce silencing of a green fluorescence protein (GFP) reporter gene. Small RNA sequencing (sRNA-seq) of the samples from dsRNA sprayed leaves revealed that the dsRNA was, if at all, not efficiently processed into siRNAs indicating that the dsRNA was insufficiently taken up by plant cells.

scholarly journals The Role of Micrornas in Genome Response to Plant–Lepidoptera Interaction

Plants ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 529
Author(s):  
Katarína Ražná ◽  
Ľudovít Cagáň
Keyword(s):  
Rna Interference ◽  
Immune Responses ◽  
Host Resistance ◽  
Defense Mechanism ◽  
Plant Protection ◽  
Regulation Of Gene Expression ◽  
Basic Principles ◽  
Double Stranded Rna ◽  
Rna Molecules

RNA interference is a known phenomenon of plant immune responses, involving the regulation of gene expression. The key components triggering the silencing of targeted sequences are double-stranded RNA molecules. The regulation of host–pathogen interactions is controlled by miRNA molecules, which regulate the expression of host resistance genes or the genes of the pathogen. The review focused on basic principles of RNA interference as a gene-silencing-based defense mechanism and the role of miRNA molecules in insect genomes. RNA interference as a tool for plant protection management is discussed. The review summarizes current miRNA-based biotechnology approaches for plant protection management.

scholarly journals Small Interfering RNA- Modern Approach for Intervening Pathological Conditions

2021 ◽  
Vol 2 (2) ◽  
pp. 16-19
Author(s):  
Shazia Choudhary ◽  
Sheeba Murad ◽  
Sana Gul ◽  
Hayat Khan ◽  
Samra Khalid ◽  
...  
Keyword(s):  
Rna Interference ◽  
Small Interfering Rna ◽  
Sirna Delivery ◽  
Rnai Pathway ◽  
Modern Approach ◽  
Rna Molecules ◽  
Rnai Technology ◽  
Efficient Delivery ◽  
Biological Discovery ◽  
Interfering Rna

RNA interference (RNAi) refers to the inhibition of gene expression by small double-stranded RNA molecules. This technology can prove to be a breakthrough biological discovery of the decade as it has the potential to revolutionize the field of therapeutics. RNA interference (RNAi) through small interfering RNA (siRNA) is currently being evaluated for its efficacy to be used in therapeutics as well as prophylactic strategies. Many studies are being conducted across the globe to optimize the siRNA delivery systems (in terms of safe, stable and efficient delivery) in various disorders. There are a number of diseases such as autoimmune diseases, cancer associated pathological changes, bacterial and viral induced disorders, where RNAi pathway can be explored and RNAi technology can be used as a tool to intervene such abnormalities. This review is an effort to review latest advancements in the field of siRNA based therapy development and the pits and falls generally encountered in the use of this technology.

SiMiSnoRNA: Collection of siRNA, miRNA, and snoRNA database for RNA interference / SiMiSnoRNA: RNA Interferansı için siRNA, miRNA ve snoRNA veritabanında depolanan siRNA, miRNA, and snoRNA koleksiyonları

2015 ◽  
Vol 40 (6) ◽  
Author(s):  
Umesh Kalathiya ◽  
Monikaben Padariya ◽  
Maciej Baginski ◽  
Chintankumar Padariya
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Small Rna ◽  
Regulation Of Gene Expression ◽  
Specific Gene ◽  
Sequence Information ◽  
Rna Molecules ◽  
Inhibit Gene Expression ◽  
Considerable Impact ◽  
Interfering Rna

AbstractObjective: The discovery of sequence specific gene silencing which occurs due to the presence of double- stranded RNAs has considerable impact on biology, revealing an unknown level of regulation of gene expression. This process is known as RNA interference (RNAi) or RNA silencing in which RNA molecules inhibit gene expression, typically by causing the destruction of specific mRNA molecule. Two types of small RNA molecules-small interfering RNA (siRNA) and microRNA (miRNA) are central to RNA interference. Therefore, SMethods: SResults: A flexible web-based search engine is developed to obtain fast access to specific small RNA sequence information.Conclusion: BLAST search analysis within S

scholarly journals Micro-RNAs

2002 ◽  
Vol 156 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Helge Grosshans ◽  
Frank J. Slack
Keyword(s):  
Gene Regulation ◽  
Rna Interference ◽  
Small Interfering Rna ◽  
Target Gene ◽  
Micro Rna ◽  
C Elegans ◽  
Micro Rnas ◽  
Key Participants ◽  
Interfering Rna ◽  
Translational Level

Two small temporally regulated RNAs (stRNAs)**Abbreviations used in this paper: stRNA, small temporally regulated RNA; miRNA, micro-RNA; siRNA, small interfering RNA; RNAi, RNA interference. of ∼22 nucleotides regulate timing of gene expression during development of the nematode C. elegans. This regulation occurs at a posttranscriptional, presumably translational, level and is distinct from RNA interference (RNAi). One of the two stRNAs, let-7, as well as its target gene, lin-41, are highly conserved even in humans, suggesting a wide employment of stRNA-mediated gene regulation. Recent reports indicate that these two stRNAs are indeed likely to represent only the tip of an iceberg with hundreds or more of additional micro-RNAs (miRNAs) existing in metazoans. miRNAs might thus be previously underestimated key participants in the field of gene regulation.

scholarly journals Dampening the silencing effect of RNA interference in mammals

2005 ◽  
Vol 390 (3) ◽  
Author(s):  
Junlong Zhang
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Target Gene ◽  
Regulation Of Gene Expression ◽  
High Dose ◽  
Double Stranded Rna ◽  
Rna Molecules ◽  
Biochemical Journal ◽  
Suppression Of Gene Expression ◽  
Interfering Rna

RNAi (RNA interference) refers to the suppression of expression of a target gene (mainly at the post-transcriptional or translational level) induced by small (21–23 nucleotides) RNA molecules, including siRNA (small interfering RNA). Suppression of gene expression by RNAi represents an important part of the regulation of gene expression. Interestingly, recent advancements in RNAi research support the notion that RNAi can be regulated just as an ordinary gene. In this issue of the Biochemical Journal, Hong et al. report their finding that suppression of RNAi is triggered by a high dose of siRNA in mice, and the suppression of RNAi in mice is related to eri-1 (enhanced RNA interference). Eri-1 is an RNaseT enzyme initially found in Caenorhabditis elegans that can degrade double-stranded RNA with 3′ overhangs. The results presented by Hong et al. have the potential to be extended and contribute to our knowledge about the regulation of RNAi in mammals.

Nanostructured Hyaluronic Acid-based Materials for the Delivery of siRNA

2018 ◽  
Vol 24 (23) ◽  
pp. 2678-2691 ◽  
Author(s):  
Keval Shah ◽  
Sunita Chawla ◽  
Anuradha Gadeval ◽  
Goutham Reddy ◽  
Rahul Maheshwari ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Rna Interference ◽  
Sirna Delivery ◽  
Treatment Strategies ◽  
Aqueous Solubility ◽  
Micro Rna ◽  
Target Space ◽  
Significant Progress ◽  
Gene Therapies ◽  
Future Potential

Background: The search for the effective treatment strategies to combat a disease that is characterized by abnormal cell growth and known as cancer is still required to reach its destiny. To address the problem, recently several gene therapies based on novel RNA interference (RNAi) have been proposed such as siRNA, micro RNA, shRNA, etc. out of which, siRNAs (silencing RNA) promises to show significant progress in pharmacotherapy, including considerable expansion of the druggable target space and the possibility of treating cancer. Methods: This review aims to uncover the hyaluronic acid (HA) and HA-hybridized nanoplatforms for siRNA delivery systems with a particular focus on the discussion of available reports while addressing the future potential of HA-based treatment strategies. Results: HA modified siRNA delivery, as promised, provided better targeting potential in many types of cancers. In addition, it was able to modify the release of siRNA as well. Toxicity of HA is well mentioned however, the loophole is yet to be filled by exploring various remedies for overcoming toxicity. Conclusion: To overcome the problems associated with these emerging genetic tools, investigators have employed glycosaminoglycan HA-based biopolymers. This biopolymer offers a variety of properties such as biodegradability, biocompatibility, aqueous solubility, viscoelasticity, and non-immunogenicity.

scholarly journals Induction of Promoter DNA Methylation Upon High-Pressure Spraying of Double-Stranded RNA in Plants

Agronomy ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 789
Author(s):  
Athanasios Dalakouras ◽  
Ioannis Ganopoulos
Keyword(s):  
High Pressure ◽  
De Novo ◽  
Epigenetic Changes ◽  
Double Stranded Rna ◽  
Rna Molecules ◽  
Promoter Sequences ◽  
Promoter Dna Methylation ◽  
Promoter Dna ◽  
First Time ◽  
De Novo Methylation

Exogenous application of RNA molecules is a potent method to trigger RNA interference (RNAi) in plants in a transgene-free manner. So far, all exogenous RNAi (exo-RNAi) applications have aimed to trigger mRNA degradation of a given target. However, the issue of concomitant epigenetic changes was never addressed. Here, we report for the first time that high-pressure spraying of dsRNAs can trigger de novo methylation of promoter sequences in plants.

scholarly journals Amphiphysin 1 Is Important for Actin Polymerization during Phagocytosis

2007 ◽  
Vol 18 (11) ◽  
pp. 4669-4680 ◽  
Author(s):  
Hiroshi Yamada ◽  
Emiko Ohashi ◽  
Tadashi Abe ◽  
Norihiro Kusumi ◽  
Shun-AI Li ◽  
...  
Keyword(s):  
Rna Interference ◽  
Sertoli Cells ◽  
Small Interfering Rna ◽  
Actin Polymerization ◽  
Actin Dynamics ◽  
Polymerization Process ◽  
Amphiphysin 1 ◽  
Phagocytic Cups ◽  
Interfering Rna ◽  
Constitutively Active

Amphiphysin 1 is involved in clathrin-mediated endocytosis. In this study, we demonstrate that amphiphysin 1 is essential for cellular phagocytosis and that it is critical for actin polymerization. Phagocytosis in Sertoli cells was induced by stimulating phosphatidylserine receptors. This stimulation led to the formation of actin-rich structures, including ruffles, phagocytic cups, and phagosomes, all of which showed an accumulation of amphiphysin 1. Knocking out amphiphysin 1 by RNA interference in the cells resulted in the reduction of ruffle formation, actin polymerization, and phagocytosis. Phagocytosis was also drastically decreased in amph 1 (−/−) Sertoli cells. In addition, phosphatidylinositol-4,5-bisphosphate–induced actin polymerization was decreased in the knockout testis cytosol. The addition of recombinant amphiphysin 1 to the cytosol restored the polymerization process. Ruffle formation in small interfering RNA-treated cells was recovered by the expression of constitutively active Rac1, suggesting that amphiphysin 1 functions upstream of the protein. These findings support that amphiphysin 1 is important in the regulation of actin dynamics and that it is required for phagocytosis.

scholarly journals Small RNA-Sequencing: Approaches and Considerations for miRNA Analysis

Diagnostics ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 964
Author(s):  
Sarka Benesova ◽  
Mikael Kubista ◽  
Lukas Valihrach
Keyword(s):  
Rna Sequencing ◽  
Small Rna ◽  
High Sensitivity ◽  
Small Rna Sequencing ◽  
Rna Seq ◽  
Liquid Biopsies ◽  
Comprehensive Overview ◽  
Rna Molecules ◽  
Novel Mirna ◽  
The Many

MicroRNAs (miRNAs) are a class of small RNA molecules that have an important regulatory role in multiple physiological and pathological processes. Their disease-specific profiles and presence in biofluids are properties that enable miRNAs to be employed as non-invasive biomarkers. In the past decades, several methods have been developed for miRNA analysis, including small RNA sequencing (RNA-seq). Small RNA-seq enables genome-wide profiling and analysis of known, as well as novel, miRNA variants. Moreover, its high sensitivity allows for profiling of low input samples such as liquid biopsies, which have now found applications in diagnostics and prognostics. Still, due to technical bias and the limited ability to capture the true miRNA representation, its potential remains unfulfilled. The introduction of many new small RNA-seq approaches that tried to minimize this bias, has led to the existence of the many small RNA-seq protocols seen today. Here, we review all current approaches to cDNA library construction used during the small RNA-seq workflow, with particular focus on their implementation in commercially available protocols. We provide an overview of each protocol and discuss their applicability. We also review recent benchmarking studies comparing each protocol’s performance and summarize the major conclusions that can be gathered from their usage. The result documents variable performance of the protocols and highlights their different applications in miRNA research. Taken together, our review provides a comprehensive overview of all the current small RNA-seq approaches, summarizes their strengths and weaknesses, and provides guidelines for their applications in miRNA research.

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