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

Författarna tackar Victoria snickare för hennes kritiska kommentarer i detta protokoll.

Del I - RNAi-medierad gen knockdown i vuxen Aedes aegypti myggor. För experimentet översikt se figur 1. 1. dsRNA Syntes <ol><li> Syntetisera specifika dsRNA mot gener av intresse och kontroll dsRNA. Observera: Vi rekommenderar u primrar för PCR-fragment i intervallet 300 till 500 baspar är belägna vid 3&#39;-änden av den specifika cDNA-2 och med den T7-primer-sekvensen bunden vid 5&#39;-änden (5&#39;-TAA TAC GAC TCA CTA TAG GG-3 &#39;). Unika fragme...

<ol>
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Det RNAi protokoll som används har utvecklats i laboratoriet Alexander Raikhel vid University of California Riverside 6,7 och liknar ett protokoll publicerats av Garver och Dimopoulos 4. Den experimentella metoden visad i denna video protokoll kan användas för att studera gener involverade i insekts diures i en in vivo-miljö. Utsöndringsorganen av insekter, de Malpighian tubuli, har väckt intresse hos generationer av forskare som en &quot;enkel&quot; modell för diures. Detta organ ?...

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

Denna video protokollet visar en effektiv teknik för att knockdown en viss gen i en insekt och genomföra en ny bioassay för att mäta utsöndringshastighet. Denna metod kan användas för att erhålla en bättre förståelse av förfarandet enligt diures hos insekter och är speciellt användbar vid studier av diures i livnär sig på blod artropoder som har förmåga att ta upp stora mängder av vätska i en enda blodmjöl. Detta RNAi-medierad gen knockdown i kombination med en in vivo-diures analys har utvecklats av Hansen labbet för att studera effekterna av RNAi-medierad knockdown av aquaporin gener på Aedes aegypti mygga diures 1. Protokollet är inställt i två delar: den första demonstrationen visar hur man konstruerar en enkel anordning mygga injektion och hur man förbereder och injicerar dsRNA i bröstkorgen av myggor för RNAi-medierad gen knockdown. Den andra demonstrationen visar hur man bestämmerutsöndringshastigheter i myggor som använder en in vivo-bioanalys.

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 detta protokoll kombinerar vi RNAi-medierad geners uttryck med en<em&gt; In vivo</em&gt; Diures analys för att studera effekterna knockdown av gener av intresse har på mygg vätskeutsöndringen.

RNAi-mediated gene Knockdown och<em&gt; In Vivo</em&gt; Diures Analys hos vuxna kvinnliga<em&gt; Aedes aegypti</em&gt; Myggor

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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The insect fat body plays a central role in insect metabolism and nutrient storage, mirroring functions of the liver and fat tissue in vertebrates. Insect ...

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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 ...