RNA Interference RNAi | siRNA RNA Interference RNAi | siRNA

What is the Difference Between the Different RNAi Methods? Hello, What is the Difference Between the Different RNAi Methods? Hvad er forskellen mellem de forskellige RNAi metoder? Hej Hvad er forskellen mellem de forskellige RNAi metoder? I mean why siRNA, shRNA, miRNA, can anyone explain the difference please? Jeg mener, hvorfor siRNA, shRNA, miRNA, kan man forklare forskellen? thanks tak
siRNA therapy side effects Dear friends, I like to disscuss about siRNA side effects, everyone that know about this can participate in this disscuss. siRNA terapi bivirkninger Kære venner, jeg gerne disscuss om siRNA bivirkninger, alle at vide om dette kan deltage i denne disscuss. For example in a orginal... For eksempel i en original ...
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Histamine H2 receptor trafficking: Role of arrestin, dynamin and clathrin in ... Related Articles Histamin H2-receptor-handel: rolle arrestin, dynamin og clathrin på ... Relaterede artikler

Histamine H2 receptor trafficking: Role of arrestin, dynamin and clathrin in H2r internalization. Histamin H2-receptor-handel: rolle arrestin, dynamin og clathrin i H2r internalisering.

Mol Pharmacol. Mol Pharmacol. 2008 Jul 10; 2008 den 10 juli;

Authors: Fernandez NC, Monczor F, Baldi A, Davio CA, Shayo C Forfattere: Fernandez NC, Monczor F, Baldi A, Davio Californien, Shayo C

Agonist-induced internalization of GPCRs has been implicated in receptor desensitization, resensitization, and downregulation. Agonist-inducerede internalisering af GPCRs har været involveret i receptor desensibilisering, resensitization, og downregulation. In the present study, we sought to establish whether the H2r agonist amthamine, besides promoting receptor desensitization, induced H2r internalization. I den foreliggende undersøgelse har vi forsøgt at fastslå, om H2r agonist amthamine, foruden at fremme receptor desensibilisering, induceret H2r internalisering. We further studied the mechanisms involved in as well as its potential role in receptor resensitization. Vi har yderligere undersøgt de mekanismer involveret i samt dets mulige rolle i receptor resensitization. In COS7 transfected cells amthamine induced H2r time-dependent internalization, showing a 70% of receptor endocytosis following 60 min exposure to amthamine. I COS7 transfected celler amthamine induceret H2r tid afhængige internalisering, der viser en 70% af receptor endocytose efter 60 min eksponering for amthamine. Agonist removal led to the rapid recovery of resensitized receptors to the cell surface. Agonist fjernelse førte til en hurtig inddrivelse af resensitized receptorer på cellens overflade. Similar results were obtained in the presence of cycloheximide, an inhibitor of protein synthesis. Lignende resultater blev opnået i nærværelse af Cicloheximid, en hæmmer af protein syntese. Treatment with okadaic acid, an inhibitor of the PP2A family of phosphatases, reduced the recovery of both H2r membrane sites and cAMP response. Behandling med okadainsyre, en hæmmer af PP2A familie af phosphatases, reduceret inddrivelse af både H2r membran websteder og Camp svar. Arrestin 3, but not arrestin 2 overexpresion reduced both H2r membrane sites and H2r-evoked cAMP response. Arrestin 3, men ikke arrestin 2 overexpresion reduceres både H2r membran websteder og H2r-evoked Camp svar. Receptor cotransfection with dominant negative mutants for arrestin, dynamin, Eps15 (component of the clathrin-mediated endocytosis machinery) or RNAi against arrestin 3 abolished both H2r internalization and resensitization. Receptor cotransfection med dominerende negative mutanter for arrestin, dynamin, Eps15 (komponent af clathrin-medieret endocytose maskiner) eller RNAi mod arrestin 3 afskaffet både H2r internalisering og resensitization. Similar results were obtained in U937 cells endogenously expressing H2r. Lignende resultater blev opnået i U937 celler endogenously udtrykke H2r. Present findings suggest that amthamine-induced H2r internalization is crucial for H2r resensitization, processes independent of H2r de novo synthesis but dependent on PP2A-mediated dephosphorylation. Nuværende resultater tyder på, at amthamine-induceret H2r internalisering er afgørende for H2r resensitization, processer uafhængige af H2r de novo syntese, men er afhængige af PP2A-medierede dephosphorylation. Although we do not provide direct evidence for H2r interaction with betaarrestin, dynamin and/or clathrin, our results support their involvement in H2r endocytosis. Selvom vi ikke giver direkte bevis for H2r interaktion med betaarrestin, dynamin og / eller clathrin, vores resultater støtte deres inddragelse i H2r endocytose. The rapid receptor recycling to the cell surface and the specific involvement of arrestin 3 in receptor internalization further suggest that the H2r belongs to class A GPCRs. Den hurtige receptor genanvendelse til cellens overflade og en specifik inddragelse af arrestin 3 i receptor-internalisering yderligere foreslå, at H2r tilhører klasse A GPCRs.

PMID: 18617631 [PubMed - as supplied by publisher] PMID: 18617631 [PubMed - som leveres af udgiveren]

A Systematic RNAi Screen Reveals Involvement of Endocytic Pathway in Neuronal Dysfunction in {alpha}-Synuclein Transgenic C. elegans. En systematisk RNAi Screen Reveals Inddragelse af Endocytic opholdet i Neuronal dysfunktion i (alpha)-Synuclein Transgene C. elegans.

Hum Mol Genet. Hum Mol Genet. 2008 Jul 9; 2008 den 9 juli;

Authors: Kuwahara T, Koyama A, Koyama S, Yoshina S, Ren CH, Kato T, Mitani S, Iwatsubo T Forfattere: Kuwahara T, Koyama A, Koyama S, Yoshina S, Ren CH, Kato T, Mitani S, Iwatsubo T

Mutations or multiplications in alpha-synuclein gene cause familial forms of Parkinson disease or dementia with Lewy bodies, and the deposition of wild-type alpha-synuclein as Lewy bodies occurs as a hallmark lesion of these disorders, collectively referred to as synucleinopathies, implicating alpha-synuclein in the pathogenesis of synucleinopathy. Mutationer eller multiplikationer i alfa-synuclein genet forårsage familiær former for Parkinsons sygdom eller demens med Lewy organer, og deposition af wild-type alfa-synuclein som Lewy organer opstår som et adelsmærke læsion af disse lidelser, kollektivt benævnt synucleinopathies, implicating alpha - synuclein i patogenese af synucleinopathy. To identify modifier genes of alpha-synuclein-induced neurotoxicity, we conducted an RNAi screen in transgenic C. elegans (Tg worms) that overexpress human alpha-synuclein in a pan-neuronal manner. At identificere ændringsord gener af alfa-synuclein-induceret neurotoksicitet, har vi foretaget en RNAi skærmen i transgene C. elegans (Tg orme), at overexpress menneskelige alpha-synuclein i et pan-neurale måde. To enhance the RNAi effect in neurons, we crossed alpha-synuclein Tg worms with an RNAi-enhanced mutant eri-1 strain. For at øge RNAi virkning i neuroner, vi krydsede alpha-synuclein Tg orme med en RNAi forbedret mutant eri-1 stamme. We tested RNAi of 1673 genes related to nervous system or synaptic functions, and identified ten genes that, upon knockdown, caused severe growth/motor abnormalities selectively in alpha-synuclein Tg worms. Vi testede RNAi af 1673 gener relateret til nervesystemet eller symptomfri funktioner, og indkredsede ti gener, der ved knockdown, forårsagede omfattende vækst / motordrevne abnormiteter selektivt i alfa-synuclein Tg orme. Among these were four genes (ie, apa-2, aps-2, eps-8 and rab-7) related to the endocytic pathway, including two subunits of AP-2 complex. Blandt disse var fire gener (dvs. apa-2, aps-2, eps-8 og RAB-7) i forbindelse med endocytic smittevej, herunder to enheder af AP-2 kompleks. Consistent with the results by RNAi, crossing alpha-synuclein Tg worms with an aps-2 mutant resulted in severe growth arrest and motor dysfunction.?alpha-Synuclein Tg worms displayed a decreased touch sensitivity upon RNAi of genes involved in synaptic vesicle endocytosis, and they also showed impaired neuromuscular transmission, suggesting that overexpression of alpha-synuclein caused a failure in uptake or recycling of synaptic vesicles. I overensstemmelse med resultaterne af RNAi, passage alpha-synuclein Tg orme med et aps-2 mutant resulteret i alvorlig vækst anholdelse og motorisk dysfunktion.? Alpha-Synuclein Tg orme vises en faldt touch følsomhed på RNAi af gener involveret i symptomfri vesicle endocytose, og de også viste nedsat neuromuskulær transmission, hvilket tyder på, at overexpression af alfa-synuclein forårsaget en fejl i optagelsen eller genanvendelse af symptomfri blærer. Furthermore, knockdown of apa-2, an AP-2 subunit, caused an accumulation of phosphorylated alpha-synuclein in neuronal cell bodies, mimicking synucleinopathy. Desuden knockdown af apa-2, en AP-2 subunit, forårsagede en ophobning af fosforyleret alpha-synuclein i neurale celle organer, efterligne synucleinopathy. Collectively, these findings raise a novel pathogenic link between endocytic pathway and alpha-synuclein-induced neurotoxicity in synucleinopathy. Kollektivt, disse resultater giver anledning til en roman patogene forbindelsen mellem endocytic læringsforløb og alfa-synuclein-induceret neurotoksicitet i synucleinopathy.

PMID: 18617532 [PubMed - as supplied by publisher] PMID: 18617532 [PubMed - som leveres af udgiveren]

Oligonucleotide sequences forming short self-complimentary hairpins can expedite the down-regulation of Coprinopsis cinerea genes. Oligonucleotide sekvenser danner korte selvstændige gratis hairpins kan fremskynde ned-regulering af Coprinopsis cinerea gener.

J Microbiol Methods. J Microbiol metoder. 2008 Jun 17; 2008 jun 17;

Authors: Costa AM, Mills PR, Bailey A, Foster GD, Challen MP Forfattere: Costa AM, Mills PR, Bailey A, Foster GD, Challen MP

Gene silencing in fungi is often induced by dsRNA hairpin forming constructs the preparation of which can require multiple cloning steps. Gene dets i svampe er ofte fremkaldt af dsRNA hairpin danne konstruktioner forberedelsen af hvilket kan kræve flere kloning trin. To simplify gene silencing in the filamentous fungi we have evaluated a high throughput cloning method for target sequences using the homobasidiomycete Coprinopsis cinerea, the GFP reporter and a commercially available vector system. For at forenkle genet dets i filamentøst svampe, vi har evalueret en høj overførselshastighed kloning metode til målet sekvenser ved hjælp af homobasidiomycete Coprinopsis cinerea, GFP reporter og en kommercielt tilgængelig vektor-system. The pSUPER RNAi Systemtrade mark, which was developed for mammalian experiments, exploits the human H1 Polymerase III (Pol III) RNA gene promoter and expedites cloning/expression of specific user-defined oligonucleotide sequences to form short self-complimentary hairpins. De pSUPER RNAi Systemtrade varemærke, som blev udviklet til pattedyr forsøg, udnytter de menneskelige H1 polymerase III (Pol III) RNA genet promotoren og expedites kloning / udtryk for en bestemt bruger-defineret oligonukleotide sekvenser til at danne korte selvstændige gratis hairpins. Transformation of C. cinerea with pSUPER constructs harboring specific oligonucleotides (19 nt stem length) enabled recovery of transformants with reduced transcripts of the GFP transgene, and were less fluorescent in protein assays and microscopic phenotypes. Transformation af C. cinerea med pSUPER konstruktioner harboring specifikke oligonucleotides (19 nt stamceller længde) aktiveret inddrivelse af transformants med nedsat udskrifter af GFP transgen, og var mindre fluorescerende i protein assays og mikroskopiske fænotyper. This technological advance should expedite functional genomic studies in C. cinerea and has wider potential for utility in other homobasidiomycete and filamentous fungi. Dette teknologiske fremskridt skal ekspedere funktionelle genomiske studier i C. cinerea og har større potentiale for nytteværdi i andre homobasidiomycete og filamentøst svampe.

PMID: 18616966 [PubMed - as supplied by publisher] PMID: 18616966 [PubMed - som leveres af udgiveren]