MonitorexosomedynamicswiththepCT-CD81-GFP(pCMV)ExosomeCyto-Tracer(alsoavailableasaCD81-RFPfusion,pCT-CD81-RFP).Providedaseitherlentivectororpre-packagedvirus,thisCyto-TracerfusesGFPtothetetraspaninCD81,whichthengetspackagedintoexosomesproducedbythetransfectedcells.GreatforcreatingstablecelllinesthatproduceGFP-labeledexosomesaswellaslabelingexosomesfromprimarycells,tumorcells,stemcells,andmore.
- Stablelentivector-basedsystem
- Idealforco-localizationstudies
- Monitorexosomedynamicsandfunctionalstudiesinrealtime
- Labelexosomesfromprimarycells,tumorcells,stemcells,andmore
Choosetheexosomequantitationmethodthat’sbestforyourstudies
| Promoter(Cat.#) | Cyto-Tracer | TargetLocation | PeptideTag |
|---|
CMV(CYTO100-PA-1/CYTO100-VA-1)
MSCV(CYTO100-PB-1/CYTO100-VB-1) | pCT-Mem-GFP | PlasmaMembrane | Neuromodulin |
CMV(CYTO101-PA-1/CYTO101-VA-1)
MSCV(CYTO101-PB-1/CYTO101-VB-1) | pCT-InnerMem-GFP | InnerLeafletofMembrane | Farnesylationsignal |
CMV(CYTO102-PA-1/CYTO102-VA-1)
MSCV(CYTO102-PB-1/CYTO102-VB-1) | pCT-Mito-GFP | Mitochondria | CytochromeCoxidaseVIII |
| CMV(CYTO103-PA-1/CYTO103-VA-1) | pCT-ER-GFP | EndoplasmicReticulum | ERtargetingconsensus |
| CMV(CYTO104-PA-1/CYTO104-VA-1) | pCT-Golgi-GFP | GolgiApparatus | Galactosyltrasferase |
| CMV(CYTO105-PA-1/CYTO105-VA-1) | pCT-Secretory-GFP | ER-Golgi Vesicles | Secretoryconsensuspeptide |
| CMV(CYTO106-PA-1/CYTO106-VA-1) | pCT-Lyso-GFP | Lysosome | CathepsinB |
| CMV(CYTO107-PA-1/CYTO107-VA-1) | pCT-Endo-GFP | Endosome | RhoB |
| CMV(CYTO107R-PA-1/CYTO107R-VA-1) | pCT-Endo-RFP | Endosome | RhoB |
CMV(CYTO108-PA-1/CYTO108-VA-1)
MSCV(CYTO108-PB-1/CYTO108-VB-1) | pCT-Pero-GFP | Peroxisome | Peroxisomalconsensussignal |
| CMV(CYTO120-PA-1/CYTO120-VA-1) | pCT-Autophago-GFP | Autophagosome | LC3 |
| CMV(CYTO120R-PA-1/CYTO120R-VA-1) | pCT-Autophago-RFP | Autophagosome | LC3 |
| CMV(CYTO109-PA-1/CYTO109-VA-1) | pCT-CD63-GFP | Exosome/Secretory | CD63Tetraspanin |
| CMV(CYTO109R-PA-1/CYTO109R-VA-1) | pCT-CD63-RFP | Exosome/Secretory | CD63Tetraspanin |
| CMV(CYTO122-PA-1/CYTO122-VA-1) | pCT-CD9-GFP | Exosome/Secretory | CD9Tetraspanin |
| CMV(CYTO123-PA-1/CYTO123-VA-1) | pCT-CD9-RFP | Exosome/Secretory | CD9Tetraspanin |
| CMV(CYTO124-PA-1/CYTO124-VA-1) | pCT-CD81-GFP | Exosome/Secretory | CD81Tetraspanin |
| CMV(CYTO125-PA-1/CYTO125-VA-1) | pCT-CD81-RFP | Exosome/Secretory | CD81Tetraspanin |
CMV(CYTO110-PA-1/CYTO110-VA-1)
MSCV(CYTO110-PB-1/CYTO110-VB-1) | pCT-Actin-GFP | Cytoskeleton | β-actin |
CMV(CYTO111-PA-1/CYTO111-VA-1)
MSCV(CYTO111-PB-1/CYTO111-VB-1) | pCT-Tublin-GFP | Cytoskeleton | α-tublin |
| CMV(CYTO111R-PA-1/CYTO111R-VA-1) | pCT-Tublin-RFP | Cytoskeleton | α-tublin |
| CMV(CYTO112-PA-1/CYTO112-VA-1) | pCT-MAP4-GFP | Microtubule | Microtubuleassociatedprotein4(MAP4) |
CMV(CYTO113-PA-1/CYTO113-VA-1)
MSCV(CYTO113-PB-1/CYTO113-VB-1) | pCT-H2B-GFP | Nucleus | H2B |
| CMV(CYTO114-PA-1/CYTO114-VA-1) | pCT-Apoptosis-Luc | Caspase3/7activation | Cyclicinactiveluciferase |
| CMV(CYTO115-PA-1/CYTO115-VA-1) | pCT-GFP-BAX | CytosoltoMitochondria | BAX |
| CMV(CYTO116-PA-1/CYTO116-VA-1) | pCT-Catenin-GFP | CytosoltoNucleus/CellMembrane | β-Cateninactivation |
| CMV(CYTO117-PA-1/CYTO117-VA-1) | pCT-CMV-PSD95-GFP | DendriteMembranes | PSD-95 |
CMV(CYTO118-PA-1/CYTO118-VA-1)
MSCV(CYTO118-PB-1/CYTO118-VB-1) | pCT-Cyto-GFP | CytosolicCompartment | GFP-Untagged |
| CMV(CYTO119-PA-1/CYTO119-VA-1) | pCT-Cyto-RFP | CytosolicCompartment | RFP-Untagged |
| MSCV(CYTO121-PB-1/CYTO121-VB-1) | pCT-Cyto-GFpLuc | GFPandLuciferaseControl | GFPandLuciferase(Untagged) |
SeesomeofourexosomeCyto-Tracersinaction
Thefollowingfigureandvideosarefrom:
GarciaNA,etal.GlucoseStarvationinCardiomyocytesEnhancesExosomeSecretionandPromotesAngiogenesisinEndothelialCells.PLoSONE.2015.10(9).PMCID:PMC4578916.
Figure4fromGarcia,etal.ExosometransferfromCMstoECs.(A)H9c2transfectedwithpCT-CD63-GFP.FACSanalysisof90%GFPpositivecells.(B)Representativeimagesfromconfocaltime-lapsemicroscopyofmouseECs(ACTB-DsRed)(EC;red)co-culturedwithH9C2-CD63-GFPcells(green),previouslyculturedfor24hin+/-Stmedium.ExosometransferfromH9C2CMstoECcanbeobserved(S1andS2Movies,below).WhitearrowsshowCD63-GFPstructuresinsideECs(C)RepresentativeimmunostainingofH9C2-CD63-GFPandHUVECco-cultures;anti-GFP(green)andanti-CD31(red).TheimagesillustrateGFPfluorescencefromCD63-GFPexosomesinCD31-positivecells(red)after24hincubationin+/-Stmedium.WhitearrowsshowCD63-GFPstructuresinsideECs.
Labeledexosomes.(Toppanels)CD63-GFPCyto-Tracerstransfectedintoahumanfibrosarcomacellline.(Bottompanels)CD9-GFPandCD9-RFPCyto-Tracersco-transfectedintoHEK293cells.
Citations:
Braganza,A,etal.(2017)UBE3BIsaCalmodulin-regulated,Mitochondrion-associatedE3UbiquitinLigase.J.Biol.Chem..2017Feb10;292(6):2470-2484.PMID:28003368
Gonzalez-King,H,etal.(2017)HypoxiaInduc
IBLeFactor-1αPotentiatesJagged1-MediatedAngiogenesisbyMesenchymalStemCell-DerivedExosomes.
StemCells.2017Jul1;35(7):1747-1759.PMID:28376567
Jawhari,S,etal.(2017)AutophagyandTrkC/NT-3signalingjoinedforcesboostthehypoxicglioblastomacellsurvival.Carcinogenesis.2017Jun1;38(6):592-603.PMID:28402394
Soares,VYR.(2017)OpapeldasvesículasextracelularesnafisiopatologiadaperdaauditivaocasionadapeloSchwannomavestibular.Thesis.;.Link:Thesis
Yoon,C,etal.(2016)DeliveryofmiR-155toretinalpigmentepithelialcellsmediatedbyBurkitt’slymphomaexosomes.TumourBiol..2016Jan1;37(1):313-21.PMID:26211004
Jawhari,S.(2015)Etudedessignalisationsautophagiqueetneurotrophiquedansdeslignéesdeglioblastomehumainactivéeslorsdel’hypoxie.Thesis.;.Link:Thesis
LoCicero,A,etal.(2015)Exosomesreleasedbykeratinocytesmodulatemelanocytepigmentation.NatCommun.2015Jun24;6:7506.PMID:26103923
VARLET,P,PU-PH,P&DJAVAHERI-MERGNY,M.(2015)SOHAJAWHARI.Thesis.;.Link:Thesis
Liang,Y.(2015)Exosome/MicrovesicleCargoContentsRecruitmentinaGlycosylation-dependentPathway.Thesis.;.Link:Thesis
Sato,Y,etal.(2014)MTCL1crosslinksandst
ABIlizesnon-centrosomalmicrotubulesontheGolgimembrane.
NatCommun.2014Nov4;5:5266.PMID:25366663
Liang,Y,etal.(2014)ComplexN-linkedglycansserveasadeterminantforexosome/microvesiclecargorecruitment.J.Biol.Chem..2014Nov21;289(47):32526-37.PMID:25261472
Fang,Q,etal.(2014)HSP90regulatesDNArepairviatheinteractionbetweenXRCC1andDNApolymeraseβ.NatCommun.2014Nov26;5:5513.PMID:25423885
相比传统的四环素病毒调控系统，SBI（system biosciences）最新推出的SparQ™ Cumate
调控系统无疑将为很多利用病毒载体做研究的实验者带来福音。这一系统通过不断的病毒转导，实
现严谨调控，强有力的反馈和可检测的表达，其主要用于诱导调控基因和microRNA的表达。这一
系统通过结合在cumate操纵子上的高亲和能力的CymR阻遏物进行调控，通过加入一种可以结合
在CymR上的无毒的小分子化合物Cumate，来解除抑制。这一系统可以反复使用，精确的开关机
制，从而实现严谨的调控，用作时空表达的研究。
图一：无渗漏调控系统原理图
相比传统的诱导系统，Cumate 具有更低的背景和强劲的表达，如图二所示，相比其他的系统，SparQ比其他系统表达强32倍，且是零渗漏。
同时SBI还可以根据您的需要定制Cumate系统！
