...electroporation studies, DsRed reporter expression was induced in ~76% of GFP+ cells, and 100% of DsRed+ cells also...mouse retina. GFP expression robustly activated the reporter gene TdTomato, whose expression was absent without.... Cepko lab researcher Jonathan Tang wanted to express a variety of proteins in single cell types in the... could use previously established mouse lines expressing GFP in specific cell types to do more than just...cells. If GFP could be co-opted to control gene expression, he could then selectively manipulate only GFP-labeled...GFP, but commonly used red fluorescent proteins dsRed, mCherry, and TdTomato do not induce transcription...and rTetR systems such that they activate gene expression from different promoters. Moving to an in vivo...

... al.) Monitor gene expression in the developing pancreas by placing FP timer DsRed-E5 under the control...fluorescent protein timers When FP timers are first expressed, they predominantly fluoresce one color but slowly...timers (whether as part of protein fusions or expressed independently) originally fluoresce green or blue...when Terskikh et al. reported the production of dsRed E5. This timer predictably transitions from green...displays predictable kinetics over 14 hours of expression in C. elegans embryos. However, this initial ...ideal FP timers: 1. Ratiometric determination of expression time The ratio of mature to immature fluorescence...fluorescence from the FP timer is dependent on the total expression time but independent of protein concentration...

...EGFP) [1, 2] or even form obligate tetramers (i.e. DsRed) [3]. This is a problem for fusion proteins. One...solution is to restrict imaging analysis to cells expressing the lowest levels of the fusion protein. This...as EGFP, but not for obligate oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can...

...in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms (E. coli, and later...and he was able to report new fluorophores like DsRed from Discosoma sp. and Katushka/mKate from Entacmaea...

.... Contains the visible marker 3xP3-DsRed. (Also known as pHD-DsRed-att). 51026 U6-BbsI-crRNA : Generates...for expression of chiRNA under the control of the Drosophila snRNA:U6:96Ab promoter. 51019 pDsRed-attP...for use in combination with tracrRNA. 51434 pHD-DsRed : vector for generating dsDNA donors for homology-directed...homology-directed repair. Contains the visible marker 3xP3-DsRed....

...transgene expression; Also expresses luciferase Lowe 9044 pMIG MSCV Plasmid for transgene expression; Also...Envelope VSV-G-expressing envelope plasmid Kasparov 35617 pCAG-Eco Envelope Ecotropic MLV expressing envelope...GFP MoMLV U6-driven plasmid for shRNA expression; Also expresses GFP; See plasmid 8452 for Puro resistance... and gene expression; Select with puromycin or screen for GFP Bartel 32702 pMSCV-loxp-dsRed-loxp-eGFP-... the rapid loss of dsRed and the activation of your gene fused to eGFP expression Clevers 18760 MSCV IRES...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA; expresses rtTA; See article for additional plasmids...pLncEXP CMV/MSV lncRNA expression plasmid Sun 60683 pLXIN-Luc MoMSV Stable expression of luciferase in mammalian...

...including: tagging genes for elucidating their expression or localization profiles, acting as a biosensor... could be taken directly from A. Victoria and expressed in any organism while still maintaining fluorescence...was the fact that it was non-toxic and could be expressed in living cells, enabling the study of dynamic...GFP derivatives, but are instead related to the dsRed protein isolated from Discosoma sp. Similar work...scientist to visualize when and where the gene is expressed. Click here to view Addgene’s collection of empty...interest can be used to effectively monitor gene expression from that promoter in a given cell type. This...and sensors at Addgene. Cell marking/selection: Expression constructs like plasmids often include GFP as...

...C-terminus of DsRed.T3. See Table 1 for a summary of Rosella’s excitation and emission spectra. DsRed is the ...monitor autophagy, the Pho8Δ60 mutant protein is expressed. Pho8Δ60 localizes to the cytosol in an inactive...yeast Rosella is targeted to the cytosol when expressed without a signal sequence and to the mitochondria...proteins: a relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic...spectra. Component Excitation Emission pH Range DsRed.T3 488, 543*, 568 nm 587 nm ~4.9 - 9 SEP ...

...of these FPs are fluorescent when ectopically expressed in most organisms. Furthermore, fusing FPs to ... The two most common uses are: 1) to test the expression level in a specific system (by measuring the ... 37°C, mCherry takes ~15min, TagRFP ~100min and DsRed ~10hours. Temperature: FPs maturation times and ... parameter is important if you are planning to express the FP in acidic environments (e.g. yeast cytosol...He is interested in everything related to gene expression, particularly at the RNA level. He maintains ...proteins and imaging technologies to study gene expression, nuclear localization, and dynamics” Curr. Opin...

... HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA expressing cells were then ... to test multiple expression systems before they achieve the appropriate expression level for their gene...create protein expression constructs fused with epitopes or fluorophores, generate expression vectors with... lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses the gRNA from a Drosophila U6:2 ...model organisms. As long as it was expressed in a cell that also expressed auxin-sensitive TIR1, any protein...cell lines expressing it. The Church lab fixed this issue by creating a new dCas9-VPR expression plasmid ...doxycycline-inducible expression. The end result is genomic integration of dCas9-VPR and controlled expression of the...

...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO shares many structural properties with DFHBI...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

...diversity Mammalian Megason 13769 pCALNL-DsRed Cre dependent DsRed expression Mammalian Cepko 13770 pCALNL-GFP...present. By expressing Cre at specific times or locations, you can precisely control expression of your gene...developed ways to tightly control Cre expression and to optimize Cre expression once it is induced. Addgene has...defines the areas in which Cre will be expressed. Cre may be expressed globally under a broadly active promoter... or expressed only in a subset of cells under a more specific promoter (e.g. Rho-Cre is expressed in the...visualization of Cre expression. Optimized Cre: Codon-optimized Cre ( iCre ) is expressed at higher levels... EGFP, and sgRNA expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral...

...pAdx-CMV-YFP 73348 Express YFP with an NLS from the CMV promoter pAdx-CMV-dsRed 73349 Expresses dsRed from the ...-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter when Cre is expressed by another vector pCDH-EF1...pCDH-EF1-DIO-tdTomato 72254 Expresses tdTomato under EF-1 promoter when Cre is expressed by another vector pCDH-CB-iCre-P2A-tdTomato-T2A-Puro...pCDH-CB-copGFP-T2A-iCre 72256 Expresses copGFP and iCre under from the CB promoter pCDH-CB-iCre 72257 Express iCre under ...pCDH-CB-FLPe-P2A-tdTomato 72259 Expresses FLPe and tdTomato from the CB promoter pCDH-EF1-Fon-copGFP 72260 Expresses copGFP from...promoter when FLP is expressed by another vector pCDH-EF1-Fon-tdTomato 72261 Expresses tdTomato from the ... promoter when FLP is expressed by another vector pCDH-EF1-FLPe 72262 Expresses FLPe from the EF-1 promoter...

...” mouse strain in which Cre expression causes the induction of both DsRed fluorescence and GFP shRNA. ...Cre-based Brainbow-expressing cells. Autobow plasmids are all-in-one versions which express self-excising ...multiple host cells in order to optimize expression. The expression plasmids for a given species are based...neural stem cells concomitant with activated DsRed2 expression. Brown et al., Genesis 2014 Jan;52(1):...Timer protein - a mutant of the fluorescent protein dsRed that changes irreversibly its color from green to...pMitoTimer from Yan’s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial...channel fluorescence images of C2C12 cells stably expressing pTRE-Tight-MitoTimer and rtTA. The cells were...

...loxP sites such that their expression is turned on or off in cells expressing Cre. The result of a system...allows the visualization of fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell...immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph nodes...efforts have been made to create transgenic mice expressing fluorescent protein reporter constructs in the...above depicts the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after...Yellow-orange fluorescent proteins like TagRFP, tdTomato, DsRed, the mKate series, or tdKatushka2 (Drobizhev et ...

...neomycin (pSIR-neo) , GFP (pSIR-GFP) , DsRed (pSIR-DsRed-Express2) , human CD2 (pSIR-hCD2) PDF 105.8 KB... sgRNA dual expression: pAC2 , pAC152 , pAC153 , pAC154 ; pmax dCas9 activator expression: pAC91 , pAC92...design and cloning All-in-one plasmid containing expression cassette for NmCas9 and BsmBI site for cloning...donor plasmid backbones pSL1180-HR-PUbECFP & pSL1180polyUBdsRED PDF 583.3 KB Zhang gRNA cloning CRISPR RNA...

...investigate protein localization, interaction, and expression. However, one caveat of FP-protein fusions (FP-chimeras...determine specific protein turnover or temporal expression using standard FP-chimeric proteins. Photoactivatable...PA-mRFP1(S146H / I161V / I197H) (3) were derived from DsRed and mRFP, respectively. When illuminated with violet...

... to Top Controlled Expression It isn’t always beneficial to constitutively express your gene of interest... on expression only at a specific time. This collection has a variety of plasmids whose expression levels... bacterial expression systems to find out more! Genome Engineering Although overexpressing or knocking...plasmids used for expression in bacterial systems Plasmid... Plasmid Collections Bacterial Expression Systems Bacterial...Bacterial Expression Systems Genome Engineering Visualization and Tagging Purification Controlled Expression...Tagging and Visualization Purification Controlled Expression Reporter Plasmids You may also like… Plasmids...

...pBad-EBFP2 - Bacterial Expression EBFP2-N1 - Mammalian Expression EBFP2-C1 - Mammalian Expression EBFP2-pBAD - ...- Mammalian Expression pCyPet-C1 - Mammalian Expression pCyPet-His - Bacterial Expression CYPet-N1 - Mammalian...pEiLOV-N1 - Mammalian Expression pGEX iLOV - Bacterial Expression iLOV-N1 - Mammalian Expression iLOV-C1 - Mammalian...Mammalian Expression pCAG-GFP - Mammalian Expression pLV-eGFP - Mammalian Lentiviral Expression Ac5-STABLE1...1 - Mammalian Expression mEGFP-N1 - Mammalian Expression mEGFP-C1 - Mammalian Expression mEGFP-pBAD - ... Mammalian Expression mTopaz-C1 - Mammalian Expression mTopaz-pBAD - Bacterial Expression mVenus 515 528... N1 - Mammalian Expression mVenus C1 - Mammalian Expression pKT0090 - Yeast Expression mVenus-pBAD - Bacterial...

...Douglas Kim , GENIE Project Calcium Soma-targeted expression of GCaMP7f or GCaMP6f Precision Calcium Imaging...Kim , GENIE Project Calcium Bicistronic vector expressing GCaMP6 and mRuby2 for imaging neural activity...2014 Jonathan Ting Calcium Intersectional viral expression of GCaMP6m/s in cells depdent on Cre, Flp, or...Karl Deisseroth , INTRSECT 2.0 Project Calcium Expression of GCaMP6f in PV cortical interneurons under ...-1636. Jordane Dimidschstein Calcium Enriched expression of GCaMP6s or GCaMP6f in axons (Constitutive ...Neurosci. 2018 Sep;21(9):1272-1280. Lin Tian Calcium Expression of GCaMP6s or GCaMP7b sensors fused to synaptophysin... Aug 4;11(1):3881. Adam Cohen Calcium GCaMP6f expression in forebrain GABA-ergic interneurons under the...