Name: rnai-mediated gene knockdown and in vivo diuresis assay in adult female aedes aegypti mosquitoes
Uploaded: 2012-07-14T14:00:00
Description: Watch this Scientific Journal Video about RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes at JoVE.com

Forfatterne takker Victoria Carpenter for hendes kritiske kommentarer i denne protokol.

Del I - RNAi-medieret gen knockdown i voksen Aedes aegypti myg. Til eksperiment oversigt se figur 1. 1. dsRNA Syntese <ol><li> Syntetisere specifikke dsRNAs mod gener af renter og kontrol dsRNAs. Bemærk: Det anbefales at udvikle primere til PCR-fragmenter i området fra 300 til 500 basepar placeret ved 3&#39;-enden af den specifikke cDNA 2 og med T7-primeren sekvensen bundet i 5&#39;-enden (5&#39;-TAA TAC GAC TCA CTA TAG GG-3 &#39;). Entydighed af de frag...

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Det RNAi anvendte protokol er blevet udviklet i laboratoriet af Alexander Raikhel ved University of California Riverside 6,7 og svarer til en protokol udgivet af Garver og Dimopoulos 4. Den eksperimentelle fremgangsmåde er vist i denne video protokol kan anvendes til at studere gener involveret i insekt diurese i en in vivo omgivelser. De ekskretionsorganerne af insekter, de Malpighian tubuli, har tiltrukket sig interesse for generationer af forskere som en &quot;simpel&quot; mod...

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rnai-mediated gene knockdown and in vivo diuresis assay in adult female aedes aegypti mosquitoes

Denne video protokol viser en effektiv teknik til at Knockdown et bestemt gen i et insekt og udføre en ny bioassay at måle udskillelse. Denne fremgangsmåde kan anvendes til at opnå en bedre forståelse af processen diurese i insekter, og er især nyttig ved undersøgelse af diurese i blod-fodring arthropoder, som er i stand til at optage store mængder væske i en enkelt blodmel. Dette RNAi-medieret gen knockdown kombineret med en in vivo-diurese assay blev udviklet af Hansen laboratoriet at undersøge virkningerne af RNAi-medieret knockdown af aquaporin gener på Aedes aegypti myg diurese 1. Protokollen er setup i to dele: den første demonstration viser, hvordan man kan konstruere en simpel myg injektionsanordning og hvordan man kan forberede og injicere dsRNA i thorax af myg for RNAi-medieret gen knockdown. Den anden demonstration viser, hvordan man fastlæggerudskillelseshastigheder i myg ved hjælp af et in vivo bioassay.

Watch this Scientific Journal Video about RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes at JoVE.com

RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes

I denne protokol vi kombinerer RNAi-medieret gen silencing med en<em&gt; In vivo</em&gt; Diurese assay til at studere virkningerne knockdown af gener af interesse har på myg fluid udskillelse.

RNAi-medieret Gene Knockdown og<em&gt; In Vivo</em&gt; Diurese Assay i Voksen Kvinde<em&gt; Aedes aegypti</em&gt; Myg

This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion ...

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RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes

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This report describes a step-by-step guide to the technique of acute intrathecal needle injections in a noninvasive manner, i.e. independent of catheter ...

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Aedes aegypti mosquitoes experimentally infected with Wolbachia are being utilized in programs to control the spread of arboviruses such as dengue, ...

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Mosquitoes represent a significant public health problem as vectors of various pathogens. For those studies that require an assessment of mosquito fitness ...