JournalofVisualizedExperimentswww.jove.comCopyright©2012JournalofVisualizedExperimentsSeptember2012|67|e4415|Page1of5VideoArticleHighefficiency,Site-specificTransfectionofAdherentCellswithsiRNAUsingMicroelectrodeArrays(MEA)ChetanPatel1,JitMuthuswamy11SchoolofBiologicalandHealthSystemsEngineering,ArizonaStateUniversityCorrespondenceto:JitMuthuswamyatjit@asu.eduURL:http://www.jove.com/video/4415DOI:doi:10.3791/4415Keywords:Bioengineering,Issue67,Genetics,MolecularBiology,BiomedicalEngineering,siRNA,transfection,electroporation,microelectrodearray,MEADatePublished:9/13/2012Citation:Patel,C.,Muthuswamy,J.Highefficiency,Site-specificTransfectionofAdherentCellswithsiRNAUsingMicroelectrodeArrays(MEA).J.Vis.Exp.(67),e4415,doi:10.3791/4415(2012).AbstractThediscoveryofRNAipathwayineukaryotesandthesubsequentdevelopmentofRNAiagents,suchassiRNAandshRNA,haveachievedapotentmethodforsilencingspecificgenes1-8forfunctionalgenomicsandtherapeutics.AmajorchallengeinvolvedinRNAibasedstudiesisthedeliveryofRNAiagentstotargetedcells.Traditionalnon-viraldeliverytechniques,suchasbulkelectroporationandchemicaltransfectionmethodsoftenlackthenecessaryspatialcontroloverdeliveryandaffordpoortransfectionefficiencies9-12.Recentadvancesinchemicaltransfectionmethodssuchascationiclipids,cationicpolymersandnanoparticleshaveresultedinhighlyenhancedtransfectionefficiencies13.However,thesetechniquesstillfailtoofferprecisespatialcontroloverdeliverythatcanimmenselybenefitminiaturizedhigh-throughputtechnologies,singlecellstudiesandinvestigationofcell-cellinteractions.Recenttechnologicaladvancesingenedeliveryhaveenabledhigh-throughputtransfectionofadherentcells14-23,amajorityofwhichusemicroscaleelectroporation.Microscaleelectroporationoffersprecisespatio-temporalcontroloverdelivery(uptosinglecells)andhasbeenshowntoachievehighefficiencies19,24-26.Additionally,electroporationbasedapproachesdonotrequireaprolongedperiodofincubation(typically4hours)withsiRNAandDNAcomplexesasnecessaryinchemicalbasedtransfectionm...
