RNA interference (RNAi) has become an important tool to study and utilize gene silencing by introducing short interfering RNA (siRNA). In order to predict the most efficient siRNAs, a new software tool, RNA Workbench (RNAWB), has been designed and is freely available (after registration) on http://www.rnaworkbench.com. In addition to the standard selection rules, RNAWB includes the possibility of statistical analyses of the applied selection rules (criteria). The role of RNA secondary structures in the RNA interference process as well as the application of sequence rules are discussed to show the applicability of the software.

• MeSH
• algoritmy * MeSH
• genový targeting metody   MeSH
• malá interferující RNA genetika   MeSH
• molekulární sekvence - údaje MeSH
• RNA interference * MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza RNA metody   MeSH
• sekvenční seřazení metody   MeSH
• software * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• malá interferující RNA MeSH

RNA interference (RNAi) has been widely applied for uncovering the biological functions of numerous genes, and has been envisaged as a pest control tool operating by disruption of essential gene expression. Although different methods, such as injection, feeding, and soaking, have been reported for successful delivery of double-stranded RNA (dsRNA), the efficiency of RNAi through oral delivery of dsRNA is highly variable among different insect groups. The German cockroach, Blattella germanica, is highly sensitive to the injection of dsRNA, as shown by many studies published previously. The present study describes a method to demonstrate that the dsRNA encapsulated with liposome carriers is sufficient to retard the degradation of dsRNA by midgut juice. Notably, the continuous feeding of dsRNA encapsulated by liposomes significantly reduces the tubulin expression in the midgut, and led to the death of cockroaches. In conclusion, the formulation and utilization of dsRNA lipoplexes, which protect dsRNA against nucleases, could be a practical use of RNAi for insect pest control in the future.

• MeSH
• dvouvláknová RNA metabolismus   MeSH
• hmyz genetika   MeSH
• liposomy metabolismus   MeSH
• RNA interference fyziologie   MeSH
• švábi genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• audiovizuální média MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• dvouvláknová RNA MeSH
• liposomy MeSH

More than a decade has passed since the discovery of RNA interference (RNAi), an eukaryotic sequence-specific degradation of mRNA induced by complementary double-stranded RNA (dsRNA). RNAi became a common tool for controlled down-regulation of gene expression in cultured cells, as well as in various model organisms. This review summarizes RNAi-based tools for silencing genes in living mammals, which include: (i) transgenic RNAi strategies, where RNAi is triggered by a transgene transmitted through the germline and (ii) approaches, where an RNAi trigger is delivered into an adult animal.

• MeSH
• genetická terapie MeSH
• geneticky modifikovaná zvířata MeSH
• genový knockdown metody   MeSH
• RNA interference * MeSH
• savci genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

BACKGROUND: RNA interference (RNAi) is a powerful approach to study a gene function. Transgenic RNAi is an adaptation of this approach where suppression of a specific gene is achieved by expression of an RNA hairpin from a transgene. In somatic cells, where a long double-stranded RNA (dsRNA) longer than 30 base-pairs can induce a sequence-independent interferon response, short hairpin RNA (shRNA) expression is used to induce RNAi. In contrast, transgenic RNAi in the oocyte routinely employs a long RNA hairpin. Transgenic RNAi based on long hairpin RNA, although robust and successful, is restricted to a few cell types, where long double-stranded RNA does not induce sequence-independent responses. Transgenic RNAi in mouse oocytes based on a shRNA offers several potential advantages, including simple cloning of the transgenic vector and an ability to use the same targeting construct in any cell type. RESULTS: Here we report our experience with shRNA-based transgenic RNAi in mouse oocytes. Despite optimal starting conditions for this experiment, we experienced several setbacks, which outweigh potential benefits of the shRNA system. First, obtaining an efficient shRNA is potentially a time-consuming and expensive task. Second, we observed that our transgene, which was based on a common commercial vector, was readily silenced in transgenic animals. CONCLUSIONS: We conclude that, the long RNA hairpin-based RNAi is more reliable and cost-effective and we recommend it as a method-of-choice when a gene is studied selectively in the oocyte.

• MeSH
• genetické vektory genetika   MeSH
• genový knockdown ekonomika   metody   MeSH
• HeLa buňky MeSH
• klonování DNA MeSH
• křížení genetické MeSH
• lidé MeSH
• malá interferující RNA metabolismus   MeSH
• myši transgenní MeSH
• myši MeSH
• oocyty metabolismus   MeSH
• plazmidy genetika   MeSH
• polymerázová řetězová reakce MeSH
• protoonkogenní proteiny c-mos genetika   metabolismus   MeSH
• RNA interference * MeSH
• transgeny genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• malá interferující RNA MeSH
• protoonkogenní proteiny c-mos MeSH

Toscana virus (TOSV) (Bunyavirales, Phenuiviridae, Phlebovirus, Toscana phlebovirus) and other related human pathogenic arboviruses are transmitted by phlebotomine sand flies. TOSV has been reported in nations bordering the Mediterranean Sea among other regions. Infection can result in febrile illness as well as meningitis and encephalitis. Understanding vector-arbovirus interactions is crucial to improving our knowledge of how arboviruses spread, and in this context, immune responses that control viral replication play a significant role. Extensive research has been conducted on mosquito vector immunity against arboviruses, with RNA interference (RNAi) and specifically the exogenous siRNA (exo-siRNA) pathway playing a critical role. However, the antiviral immunity of phlebotomine sand flies is less well understood. Here we were able to show that the exo-siRNA pathway is active in a Phlebotomus papatasi-derived cell line. Following TOSV infection, distinctive 21 nucleotide virus-derived small interfering RNAs (vsiRNAs) were detected. We also identified the exo-siRNA effector Ago2 in this cell line, and silencing its expression rendered the exo-siRNA pathway largely inactive. Thus, our data show that this pathway is active as an antiviral response against a sand fly transmitted bunyavirus, TOSV.

• MeSH
• arboviry * genetika   MeSH
• lidé MeSH
• malá interferující RNA genetika   MeSH
• Phlebotomus * genetika   MeSH
• Phlebovirus * genetika   MeSH
• Psychodidae * genetika   MeSH
• RNA interference MeSH
• virus horečky pappataci * genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• malá interferující RNA MeSH

Mitochondrial RNA-binding proteins MRP1 and MRP2 occur in a heteromeric complex that appears to play a role in U-insertion/deletion editing in trypanosomes. Reduction in the levels of MRP1 (gBP21) and/or MRP2 (gBP25) mRNA by RNA interference in procyclic Trypanosoma brucei resulted in severe growth inhibition. It also resulted in the loss of both proteins, even when only one of the MRP mRNAs was reduced, indicating a mutual dependence for stability. Elimination of the MRPs gave rise to substantially reduced levels of edited CyB and RPS12 mRNAs but little or no reduction of the level of edited Cox2, Cox3, and A6 mRNAs as measured by poisoned primer extension analyses. In contrast, edited NADH-dehydrogenase (ND) subunit 7 mRNA was increased 5-fold in MRP1+2 double knock-down cells. Furthermore, MRP elimination resulted in reduced levels of Cox1, ND4, and ND5 mRNAs, which are never edited, whereas mitoribosomal 12 S rRNA levels were not affected. These data indicate that MRP1 and MRP2 are not essential for RNA editing per se but, rather, play a regulatory role in the editing of specific transcripts and other RNA processing activities.

• MeSH
• časové faktory MeSH
• DNA primery chemie   MeSH
• editace RNA MeSH
• glycerol chemie   MeSH
• imunoprecipitace MeSH
• klonování DNA MeSH
• messenger RNA metabolismus   MeSH
• mitochondriální proteiny metabolismus   fyziologie   MeSH
• northern blotting MeSH
• plazmidy metabolismus   MeSH
• proteiny vázající RNA metabolismus   fyziologie   MeSH
• protozoální proteiny metabolismus   fyziologie   MeSH
• RNA interference * MeSH
• RNA protozoální MeSH
• RNA ribozomální metabolismus   MeSH
• RNA chemie   metabolismus   MeSH
• Southernův blotting MeSH
• transfekce MeSH
• Trypanosoma brucei brucei metabolismus   MeSH
• vazba proteinů MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Názvy látek
• DNA primery MeSH
• gBP21 protein, Trypanosoma brucei MeSH Prohlížeč
• gBP25 protein, Trypanosoma brucei MeSH Prohlížeč
• glycerol MeSH
• messenger RNA MeSH
• mitochondriální proteiny MeSH
• proteiny vázající RNA MeSH
• protozoální proteiny MeSH
• RNA protozoální MeSH
• RNA ribozomální MeSH
• RNA, ribosomal, 12S MeSH Prohlížeč
• RNA MeSH

In mammals, RNA interference (RNAi) was historically studied as a cytoplasmic event; however, in the last decade, a growing number of reports convincingly show the nuclear localization of the Argonaute (AGO) proteins. Nevertheless, the extent of nuclear RNAi and its implication in biological mechanisms remain to be elucidated. We found that reduced Lamin A levels significantly induce nuclear influx of AGO2 in SHSY5Y neuroblastoma and A375 melanoma cancer cell lines, which normally have no nuclear AGO2. Lamin A KO manifested a more pronounced effect in SHSY5Y cells compared to A375 cells, evident by changes in cell morphology, increased cell proliferation, and oncogenic miRNA expression. Moreover, AGO fPAR-CLIP in Lamin A KO SHSY5Y cells revealed significantly reduced RNAi activity. Further exploration of the nuclear AGO interactome by mass spectrometry identified FAM120A, an RNA-binding protein and known interactor of AGO2. Subsequent FAM120A fPAR-CLIP, revealed that FAM120A co-binds AGO targets and that this competition reduces the RNAi activity. Therefore, loss of Lamin A triggers nuclear AGO2 translocation, FAM120A mediated RNAi impairment, and upregulation of oncogenic miRNAs, facilitating cancer cell proliferation.

• MeSH
• aktivní transport - buněčné jádro MeSH
• Argonaut proteiny * metabolismus   genetika   MeSH
• buněčné jádro * metabolismus   MeSH
• lamin typ A * metabolismus   genetika   MeSH
• lidé MeSH
• melanom genetika   metabolismus   patologie   MeSH
• mikro RNA * metabolismus   genetika   MeSH
• nádorové buněčné linie MeSH
• proliferace buněk * genetika   MeSH
• proteiny vázající RNA metabolismus   genetika   MeSH
• RNA interference * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• AGO2 protein, human MeSH Prohlížeč
• Argonaut proteiny * MeSH
• lamin typ A * MeSH
• mikro RNA * MeSH
• proteiny vázající RNA MeSH

ADAR RNA editing enzymes (adenosine deaminases acting on RNA) that convert adenosine bases to inosines were first identified biochemically 30 years ago. Since then, studies on ADARs in genetic model organisms, and evolutionary comparisons between them, continue to reveal a surprising range of pleiotropic biological effects of ADARs. This review focuses on Drosophila melanogaster, which has a single Adar gene encoding a homolog of vertebrate ADAR2 that site-specifically edits hundreds of transcripts to change individual codons in ion channel subunits and membrane and cytoskeletal proteins. Drosophila ADAR is involved in the control of neuronal excitability and neurodegeneration and, intriguingly, in the control of neuronal plasticity and sleep. Drosophila ADAR also interacts strongly with RNA interference, a key antiviral defense mechanism in invertebrates. Recent crystal structures of human ADAR2 deaminase domain-RNA complexes help to interpret available information on Drosophila ADAR isoforms and on the evolution of ADARs from tRNA deaminase ADAT proteins. ADAR RNA editing is a paradigm for the now rapidly expanding range of RNA modifications in mRNAs and ncRNAs. Even with recent progress, much remains to be understood about these groundbreaking ADAR RNA modification systems.

• Klíčová slova
• ADAR, Drosophila melanogaster, RNA editing, RNA modification, dsRNA, epitranscriptome,
• MeSH
• adenosindeaminasa chemie   genetika   metabolismus   MeSH
• Drosophila melanogaster genetika   metabolismus   MeSH
• editace RNA * MeSH
• exprese genu MeSH
• interakční proteinové domény a motivy MeSH
• izoenzymy MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• molekulární evoluce MeSH
• nervový systém metabolismus   MeSH
• obratlovci MeSH
• proteiny Drosophily genetika   metabolismus   MeSH
• proteiny vázající RNA genetika   metabolismus   MeSH
• RNA interference MeSH
• substrátová specifita MeSH
• vazba proteinů MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• Adar protein, Drosophila MeSH Prohlížeč
• adenosindeaminasa MeSH
• izoenzymy MeSH
• messenger RNA MeSH
• proteiny Drosophily MeSH
• proteiny vázající RNA MeSH

Throughout eukaryotes, the gene encoding subunit 6 (ATP6) of the F(1)F(O)-ATP synthase (complex V) is maintained in mitochondrial (mt) genomes, presumably because of its high hydrophobicity due to its incorporation into the membrane-bound F(O) moiety. In Trypanosoma species, a mt transcript that undergoes extensive processing by RNA editing has a very low sequence similarity to ATP6 from other organisms. The notion that the putative ATP6 subunit is assembled into the F(O) sub-complex is ostensibly challenged by the existence of naturally occurring dyskinetoplastic (Dk) and akinetoplastid (Ak) trypanosomes, which are viable despite lacking the mtDNA required for its expression. Taking advantage of the different phenotypes between RNA interference knock-down cell lines in which the expression of proteins involved in mtRNA metabolism and editing can be silenced, we provide support for the view that ATP6 is encoded in the mt genome of Trypanosoma species and that it is incorporated into complex V. The reduction of the F(1)F(O) oligomer of complex V coincides with the accumulation of the F(1) moiety in ATP6-lacking cells, which also appear to lack the F(O) ATP9 multimeric ring. The oligomycin sensitivity of ATPase activity of complex V in ATP6-lacking cells is reduced, reflecting the insensitivity of the Dk and Ak cells to this drug. In addition, the F(1) moiety of complex V appears to exist as a dimer in steady state conditions and contains the ATP4 subunit traditionally assigned to the F(O) sub-complex.

• MeSH
• editace RNA * MeSH
• inhibitory enzymů farmakologie   MeSH
• membránové potenciály MeSH
• mitochondrie genetika   metabolismus   MeSH
• oligomyciny farmakologie   MeSH
• protonové ATPasy antagonisté a inhibitory   genetika   metabolismus   MeSH
• protozoální proteiny genetika   metabolismus   MeSH
• RNA interference * MeSH
• Trypanosoma brucei brucei účinky léků   enzymologie   genetika   růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• inhibitory enzymů MeSH
• oligomyciny MeSH
• protonové ATPasy MeSH
• protozoální proteiny MeSH

Since many people all around the world are suffering from genetic disorders, modern therapeutic approaches are focused on the search of new pharmaceutical products. These products will be able to act on the gene level, more accurately on the nucleotide sequences themselves. RNA interference (RNAi) is an evolutionary conserved process that is caused by double stranded RNA (dsRNA). MicroRNA (miRNA) and small interfering RNA (siRNA) are the most important dsRNAs, which have been identified so far. Short (19-25 bp) non-coding dsRNAs are responsible for regulation of cellular development, heterochromatin formation and genomic stability in eukaryotes. Most importantly they are able to silence cognate genes. Therefore, they can provide new insights into the gene function and pathway analysis. Furthermore, they are believed to be new potential targets for diagnosis and therapeutics, especially for the treatment of genetic disorders, which can be caused by nucleotides insertions, deletions and translocations.

• MeSH
• genetická terapie MeSH
• lidé MeSH
• malá interferující RNA fyziologie   terapeutické užití   MeSH
• mikro RNA fyziologie   terapeutické užití   MeSH
• RNA interference fyziologie   MeSH
• umlčování genů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• malá interferující RNA MeSH
• mikro RNA MeSH