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https://www.readbyqxmd.com/read/28816598/short-versus-long-double-stranded-rna-activation-of-a-post-transcriptional-gene-knockdown-pathway
#1
Nir Shpak, Rivka Manor, Lihie Katzir Abilevich, Ortal Mantal, Keshet Shavit, Eliahu D Aflalo, Debra Toiber, Amir Sagi
RNA interference (RNAi) utilizes a conserved cellular autoimmune defense mechanism involving the internalization of dsRNA into cells and the activation of a set of RNAi related genes. Using RNAi, complete sex reversal is achievable in males of the prawn Macrobrachium rosenbergii by knocking down the transcript level of an insulin-like androgenic gland hormone (Mr-IAG) through injections of dsRNA of the entire Mr-IAG ORF sequence (dsMr-IAG - 518 bp). Interestingly, in-vivo knockdown success and dsMr-IAG lengths seemed to correlate, with long dsRNA being the most effective and short dsRNA fragments showing no effect...
August 17, 2017: RNA Biology
https://www.readbyqxmd.com/read/28815217/deletion-of-cytoplasmic-double-stranded-rna-sensors-does-not-uncover-viral-small-interfering-rna-production-in-human-cells
#2
Susan Schuster, Lotte E Tholen, Gijs J Overheul, Frank J M van Kuppeveld, Ronald P van Rij
Antiviral immunity in insects and plants is mediated by the RNA interference (RNAi) pathway in which viral long double-stranded RNA (dsRNA) is processed into small interfering RNAs (siRNAs) by Dicer enzymes. Although this pathway is evolutionarily conserved, its involvement in antiviral defense in mammals is the subject of debate. In vertebrates, recognition of viral RNA induces a sophisticated type I interferon (IFN)-based immune response, and it has been proposed that this response masks or inhibits antiviral RNAi...
July 2017: MSphere
https://www.readbyqxmd.com/read/28811334/argonaute-8-ago8-mediates-the-elicitation-of-primary-defense-against-herbivory
#3
Maitree Pradhan, Priyanka Pandey, Klaus Gase, Murali Shraff, Ravi K Singh, Avinash Sethi, Ian T Baldwin, Shree P Pandey
In Nicotiana attenuata, specific RNA directed RNA polymerase (RdR1) and the Dicer-like proteins (DCL3 and 4) are recruited during herbivore attack to mediate the regulation of defense responses. However, the identity and role(s) of Argonautes (AGOs) involved in herbivory remains unknown. Of the 11 AGOs in the N. attenuata genome, we silenced the expression of 10. Plants silenced in NaAGO8 expression grew normally but were highly susceptible to herbivore attack. Larvae of the Manduca sexta grew faster when consuming irAGO8 plants, but did not differ from WT when consuming plants silenced in AGO1 (a, b and c), AGO2, AGO4 (a and b), AGO7, or AGO10 expression...
August 15, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28809941/analysis-of-agoshrna-maturation-and-loading-into-ago2
#4
Alex Harwig, Zita Kruize, Zhenhuang Yang, Tobias Restle, Ben Berkhout
The RNA interference (RNAi) pathway was recently expanded by the discovery of multiple alternative pathways for processing of natural microRNA (miRNA) and man-made short hairpin RNA (shRNA) molecules. One non-canonical pathway bypasses Dicer cleavage and requires instead processing by Argonaute2 (Ago2), which also executes the subsequent silencing step. We named these molecules AgoshRNA, which generate only a single active RNA strand and thus avoid off-target effects that can be induced by the passenger strand of a regular shRNA...
2017: PloS One
https://www.readbyqxmd.com/read/28805496/natural-antisense-transcripts-drive-a-regulatory-cascade-controlling-c-myc-transcription
#5
Sara Napoli, Valentina Piccinelli, Sarah Mapelli, Giuseppina Pisignano, Carlo V Catapano
Cis-natural antisense transcripts (cis-NATs) are long noncoding RNAs transcribed from the opposite strand and overlapping coding and noncoding genes on the sense strand. cis-NATs are widely present in the human genome and can be involved in multiple mechanisms of gene regulation. Here, we describe the presence of cis-NATs in the 3' distal region of the c-MYC locus and investigate their impact on transcriptional regulation of this key oncogene in human cancers. We found that cis-NATs are produced as consequence of the activation of cryptic transcription initiation sites in the 3' distal region downstream of the c-MYC 3'UTR...
August 14, 2017: RNA Biology
https://www.readbyqxmd.com/read/28796998/microrna-dependent-control-of-serotonin-induced-pulmonary-arterial-contraction
#6
Diana Dahan, Tran Thi Hien, Philip Tannenberg, Mari Ekman, Catarina Rippe, Thomas Boettger, Thomas Braun, Karin Tran-Lundmark, Phan-Kiet Tran, Karl Swärd, Sebastian Albinsson
BACKGROUND: Serotonin (5-HT) is considered to play a role in pulmonary arterial hypertension by regulating vascular remodeling and smooth muscle contractility. Here, arteries from mice with inducible and smooth muscle-specific deletion of Dicer were used to address mechanisms by which microRNAs control 5-HT-induced contraction. METHODS: Mice were used 5 weeks after Dicer deletion, and pulmonary artery contractility was analyzed by wire myography. RESULTS: No change was seen in right ventricular systolic pressure following dicer deletion, but systemic blood pressure was reduced...
August 11, 2017: Journal of Vascular Research
https://www.readbyqxmd.com/read/28793796/platelet-enriched-micrornas-and-cardiovascular-homeostasis
#7
Amro Elgheznawy, Ingrid Fleming
SIGNIFICANCE: Platelets are anucleate blood cells involved in hemostasis and thrombosis. Although no longer able to generate RNA de novo, platelets contain mRNA, YRNA fragments and premature microRNAs (miRNAs) that inherit from megakaryocytes. Recent Advances. Novel sequencing techniques have helped identify the unexpectedly large number of RNA species present in platelets. Throughout their life time platelets can process the pre-existing pool of premature miRNA to give the fully functional miRNA that can regulated platelet protein expression and function...
August 10, 2017: Antioxidants & Redox Signaling
https://www.readbyqxmd.com/read/28789966/pecanex-functions-as-a-competitive-endogenous-rna-of-s-phase-kinase-associated-protein-2-in-lung-cancer
#8
Jingqiu Li, Haihua Tian, Jinchang Pan, Nan Jiang, Jie Yang, Chengwei Zhou, Dazhi Xu, Xiaodan Meng, Zhaohui Gong
Investigating the RNA-RNA interactions involving in the initiation and progression of non-small cell lung cancer (NSCLC) may provide promising diagnostic and targeted therapeutic strategies. Here, we showed that pecanex (PCNX) positively regulates the mRNA and protein expressions of S-phase kinase associated protein 2 (Skp2) in miRNA- and 3' UTR-dependent manners. And miR-26, miR-182, miR-340 and miR-506 were verified as the common miRNAs shared by Skp2 and PCNX. Intriguingly, we initially uncovered that PCNX-3' UTR promotes cell growth, proliferation and cell cycle progression, and suppresses apoptosis of lung cancer cells, which is consistent with the oncogenic activity of Skp2-3' UTR...
August 5, 2017: Cancer Letters
https://www.readbyqxmd.com/read/28777373/the-mir-183-itga3-axis-is-a-key-regulator-of-prosensory-area-during-early-inner-ear-development
#9
Priscilla Van den Ackerveken, Anaïs Mounier, Aurelia Huyghe, Rosalie Sacheli, Pierre-Bernard Vanlerberghe, Marie-Laure Volvert, Laurence Delacroix, Laurent Nguyen, Brigitte Malgrange
MicroRNAs are important regulators of gene expression and are involved in cellular processes such as proliferation or differentiation, particularly during development of numerous organs including the inner ear. However, it remains unknown if miRNAs are required during the earliest stages of otocyst and cochlear duct development. Here, we report that a conditional loss of Dicer expression in the otocyst impairs the early development of the inner ear as a result of the accumulation of DNA damage that trigger p53-mediated apoptosis...
August 4, 2017: Cell Death and Differentiation
https://www.readbyqxmd.com/read/28769895/micrornas-as-important-players-in-host-adenovirus-interactions
#10
REVIEW
Diogo Piedade, José M Azevedo-Pereira
MicroRNAs (miRNAs) are powerful regulators of gene expression and fine-tuning genes in all tissues. Cellular miRNAs can control 100s of biologic processes (e.g., morphogenesis of embryonic structures, differentiation of tissue-specific cells, and metabolic control in specific cell types) and have been involved in the regulation of nearly all cellular pathways. Inherently to their involvement in different physiologic processes, miRNAs deregulation has been associated with several diseases. Moreover, several viruses have been described as either, avoid and block cellular miRNAs or synthesize their own miRNA to facilitate infection and pathogenesis...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28754532/centenarians-maintain-mirna-biogenesis-pathway-while-it-is-impaired-in-octogenarians
#11
C Borrás, E Serna, J Gambini, M Inglés, J Vina
Centenarians but not octogenarians up regulate the expression of miRNAs, as we previously reported. We have looked into miRNA biogenesis. We show that RNA POL II, DROSHA, EXPORTIN 5 and DICER, are up-regulated in centenarians compared with octogenarians. Furthermore, factors involved in the control of these miRNAs biogenesis genes are also up-regulated in centenarians. Therefore, the up-regulation of miRNA expression in centenarians can be explained in part because miRNA biogenesis pathway is depressed in octogenarians (ordinary aging) while it is maintained in centenarians (extraordinary aging)...
July 25, 2017: Mechanisms of Ageing and Development
https://www.readbyqxmd.com/read/28753441/rnai-mediated-resistance-to-viruses-a-critical-assessment-of-methodologies
#12
REVIEW
Mikhail M Pooggin
In plants, RNA interference (RNAi)-based antiviral defense is mediated by multigenic families of Dicer-like enzymes generating small interfering (si)RNAs from double-stranded RNA (dsRNA) produced during replication and/or transcription of RNA and DNA viruses, and Argonaute enzymes binding viral siRNAs and targeting viral RNA and DNA for siRNA-directed posttranscriptional and transcriptional silencing. Successful viruses are able to suppress or evade the production or action of viral siRNAs. In antiviral biotech approaches based on RNAi, transgenic expression or non-transgenic delivery of dsRNA cognate to a target virus pre-activates or boosts the natural plant antiviral defenses...
July 25, 2017: Current Opinion in Virology
https://www.readbyqxmd.com/read/28743868/virus-and-dsrna-triggered-transcriptional-responses-reveal-key-components-of-honey-bee-antiviral-defense
#13
Laura M Brutscher, Katie F Daughenbaugh, Michelle L Flenniken
Recent high annual losses of honey bee colonies are associated with many factors, including RNA virus infections. Honey bee antiviral responses include RNA interference and immune pathway activation, but their relative roles in antiviral defense are not well understood. To better characterize the mechanism(s) of honey bee antiviral defense, bees were infected with a model virus in the presence or absence of dsRNA, a virus associated molecular pattern. Regardless of sequence specificity, dsRNA reduced virus abundance...
July 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28739418/deficiency-of-dgcr8-increases-bone-formation-through-downregulation-of-mir-22-expression
#14
Young-Jin Choi, Sujin Jeong, Kyung-Ae Yoon, Hye-Jin Sung, Hye-Sim Cho, Dong Wook Kim, Je-Yoel Cho
MicroRNAs (miRNA) significantly contribute to bone formation by post-transcriptional regulation of gene expression. Mature miRNAs are generated following sequential cleavage by DROSHA/DGCR8 and DICER. However, recent studies have identified that some miRNAs require only one of these enzymes. Most studies seeking to clarify the role of miRNA during bone formation have been performed using DICER deletion strategies, but little is known regarding the role of DGCR8. To study the function of DGCR8 in osteogenesis, we generated mice in which Dgcr8 is conditionally deleted in osteoprogenitor cells by Col1a1-Cre...
July 21, 2017: Bone
https://www.readbyqxmd.com/read/28730448/preparation-of-a-conditional-rna-switch
#15
Paul Zakrevsky, Lorena Parlea, Mathias Viard, Eckart Bindewald, Kirill A Afonin, Bruce A Shapiro
RNA has gained great interest for use in biomedical and therapeutic applications. This is due in part to RNA's ability to perform multiple functions, including the regulation of endogenously expressed genes. However, the ability of RNA based drugs to distinguish target diseased cells from healthy tissue remains challenging. Here we present methods for the production of a recently developed conditional RNA switch that releases a Dicer substrate RNA in response to interaction with a specific RNA biomarker.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28730440/cellular-delivery-of-sirnas-using-bolaamphiphiles
#16
Kshitij Gupta
Discovery of RNA interference (RNAi) has opened up a new arena of therapeutic intervention for the treatment of cancerous as well as noncancerous diseases. The RNAi pathway utilizes RNAi inducers such as small interfering RNAs (siRNAs) to target and silence disease causing genes. However, efficient delivery of siRNAs for eliciting efficacious RNAi has remained a daunting challenge. Nonviral vectors such as lipids have shown great promise in delivering siRNAs. Recently, a novel class of cationic lipid molecules "bolaamphiphile lipids" or "bola lipids" has been shown to deliver siRNAs to cause effective gene silencing in cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28729028/micrornas-as-key-effectors-in-the-p53-network
#17
Frauke Goeman, Sabrina Strano, Giovanni Blandino
The guardian of the genome p53 is embedded in a fine-spun network of MicroRNAs. p53 is able to activate or repress directly the transcription of MicroRNAs that are participating in the tumor-suppressive mission of p53. On the other hand, the expression of p53 is under tight control of MicroRNAs that are either targeting directly p53 or factors that are modifying its protein level or activity. Although the most important function of p53 is suggested to be transcriptional regulation, there are several nontranscriptional functions described...
2017: International Review of Cell and Molecular Biology
https://www.readbyqxmd.com/read/28723582/overexpression-and-purification-of-dicer-and-accessory-proteins-for-biochemical-and-structural-studies
#18
Niladri K Sinha, Brenda L Bass
The Dicer family of ribonucleases plays a key role in small RNA-based regulatory pathways by generating short dsRNA fragments that modulate expression of endogenous genes, or protect the host from invasive nucleic acids. Beginning with its initial discovery, biochemical characterization of Dicer has provided insight about its catalytic properties. However, a comprehensive understanding of how Dicer's domains contribute to substrate-specific recognition and catalysis is lacking. One reason for this void is the lack of high-resolution structural information for a metazoan Dicer in the apo- or substrate-bound state...
July 16, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28717424/low-active-loading-of-cargo-into-engineered-extracellular-vesicles-results-in-inefficient-mirna-mimic-delivery
#19
Dhruvitkumar S Sutaria, Jinmai Jiang, Ola A Elgamal, Steven M Pomeroy, Mohamed Badawi, Xiaohua Zhu, Ryan Pavlovicz, Ana Clara P Azevedo-Pouly, Jeffrey Chalmers, Chenglong Li, Mitch A Phelps, Thomas D Schmittgen
Extracellular vesicles (EVs) hold great potential as novel systems for nucleic acid delivery due to their natural composition. Our goal was to load EVs with microRNA that are synthesized by the cells that produce the EVs. HEK293T cells were engineered to produce EVs expressing a lysosomal associated membrane, Lamp2a fusion protein. The gene encoding pre-miR-199a was inserted into an artificial intron of the Lamp2a fusion protein. The TAT peptide/HIV-1 transactivation response (TAR) RNA interacting peptide was exploited to enhance the EV loading of the pre-miR-199a containing a modified TAR RNA loop...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28716689/brg1-and-smarcal1-transcriptionally-co-regulate-drosha-dgcr8-and-dicer-in-response-to-doxorubicin-induced-dna-damage
#20
Ketki Patne, Radhakrishnan Rakesh, Vijendra Arya, Upasana Bedi Chanana, Ramesh Sethy, Pynskhem Bok Swer, Rohini Muthuswami
Recent investigations have emphasized the role of miRNA biogenesis proteins in the synthesis of non-coding RNA when double-strand DNA breaks are induced by ionizing radiations. However, the role of these non-coding RNA and their regulation in response to doxorubicin-induced DNA damage is not known. In this paper, BRG1 and SMARCAL1, members of the ATP-dependent chromatin remodelling family, are shown to co-regulate the transcription of DROSHA, DGCR8, and DICER in response to double-strand DNA breaks induced by doxorubicin...
July 15, 2017: Biochimica et Biophysica Acta
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