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We narrowed to 22 results for: TOP-GFP reporter element

Showing: 1 - 20 of 22 results

  1. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    Published
    June 2, 2020, 1:15 p.m.
    ...https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed...calcineurin transcriptional reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X...bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate... Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial...selectively remove a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is ...in turn binds to calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin... it is an attractive drug target. Calcineurin reporters allow scientists to measure calcineurin dynamics...

  2. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ... 2, the cassette should contain a reporter DNA sequence (such as GFP) in the sense orientation, followed...invert the reporter and GeneR fragment. Then the second recombination event would excise the reporter from ...Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered using AAV injection...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ...Cre-dependent reporter expression, the potential for background expression of the reporter will be greatly...-the FLEx switch--which utilizes recombination elements to turn off expression of one gene, while simultaneously...sequence in the antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by two...

  3. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as... example, if green fluorescent protein (GFP) was the reporter gene and green cells were observed in the... screens to introduce reporters throughout an organism’s genome. These reporters have historically been...stimulate the expression of the reporter. The system is made up of a reporter gene flanked by a minimal promoter...promoter – all rely on the random integration of a reporter construct in close proximity to an endogenous ...developmental stage you could infer that the gene which GFP was monitoring was being expressed in the same pattern... for with an uncharacterized gene. The actual reporter construct can be something phenotypic (such as...

  4. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown...gene editing. Dr. Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy...identified a family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on...mutagenesis libraries or to insert heterologous genes, reporters, or other markers into a target genome. They did... did this by selecting the important elements from existing transposon and vector systems and creating...an all-synthetic vector that included only the elements needed for function. The lab validated this vector...

  5. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...molecule NSDB and Na+ hypertonicity are key elements of iTOP. iTOP can be used to transduce Cas9 and a gRNA... Neuronal stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo.... Find the Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery Techniques... cells and activate expression of a luciferase reporter. CPP was not required for protein entry into cells...primary cell types using Cre and various loxP reporters. In both mouse and human embryonic stem cells,...studies showed that iTOP functions through macropinocytosis. D’Astolfo et al. tested iTOP in a series of ... transduction, iTOP is at least four times more efficient in primary fibroblasts. iTOP also works in multiple...

  6. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ... are key elements of an AAV plasmid. Parts shown in grey are optional cis-regulatory elements.   Key....4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such as ...sequences. Other cis-regulatory elements Next let’s talk about some of the elements that aren’t required parts... common AAV plasmid elements! And while there are many other AAV plasmid elements and combinations out...Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...AAV transfer plasmids have a promoter, but this element isn’t required when the transfer plasmid harbors...Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when transcribed...

  7. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...cloning in genome target sequence into EGFP reporter. Target DNA reporter system: Can evaluate component delivery...Do you want to create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively...is typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells ...separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells....compatible with SaCas9 (packaging limit ~4.5kb). CRISPR elements are inserted into an AAV transfer vector and used...published as CRISPR testing tools: Traffic Light Reporter System: Can evaluate lentiviral component delivery...repair by non-homologous end joining (NHEJ) or HDR. EGFP validation of sgRNAs: Can evaluate component delivery...

  8. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...Map of a plasmid with its elements described below.   Vector Element Description Origin of Replication... about a specific plasmid element that you would like answered or any topic suggestions for our Plasmids...expression plasmids, gene knock-down plasmids, reporter plasmids, viral plasmids, and genome engineering...cell Plasmids 101: Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of...the fundamentals. Our mission is to curate a one-stop reference guide for plasmids, so that you can spend...

  9. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ...two genes at a time using IRES elements. Further, scientists have reported lower expression of the downstream...plasmid that already has the multicistronic element and reporter cloned in. In these plasmids you would simply...stream of the IRES or 2A element (depending on the placement of the reporter gene). Addgene's collection...flanking the multicistronic element(s), or you could replace one of the reporter genes above with your gene... mutually exclusive elements. Labs have successfully utilized 2A and IRES elements within the same multicistronic... Plasmid Name Multicistronic Element Expression Type MSCV-IRES-EGFP IRES Retroviral pMSCV-pBabeMCS-IRES-RFP...

  10. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...about Mammalian Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org Find ...it's time to take a look at some of its parts. Elements such as the antibiotic resistance gene and the...the plasmid express our gene of interest. This element is responsible for initiating the transcription...polymerase binding site) and operators (response elements). Promoters are about 100 to 1000 base pairs long...RNAP can recognize and bind to specific promoter elements. This means that the promoter present in your ... other regulatory factors: enhancers, boundary elements, insulators, and silencers; however, some "leakiness...typically include some combination of other promoter elements, and tend to be more tightly regulated. Common...

  11. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ... intensity-based Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren ...line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique...and labeled with primary antibody (0.01 µg/mL Anti-GFP [N86/38.1R] (Addgene #180084), 0.01 µg/mL Anti-HA...based on several new site-specific retrotransposon elements they identified and characterized (Fell et al....untranslated regions (UTRs) outside of the homology elements allowed for scarless insertion — no UTR sequence...protein labeling compared to immunoprecipitation. The top panel depicts the identification of heat-shock proteins...optogenetic activator channelrhodopsin-2 or fluorescent reporter dTomato under the control of one of seven enhancers...

  12. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ... containing TetO elements (often linked in groups of seven as a Tet Response Element (TRE)), allowing ...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...), TetR preferentially binds Tet over the TetO elements, allowing transcription to proceed (pJKR-H-tetR...Church lab uses this system to control expression of GFP.)  To turn this inducible system into a repressible...component, the GAL80 repressor, adds a repressible element to this binary system (making this a repressible...2006): 703-9. PubMed PMID: 16582903. Potter, Christopher J., et al. “The Q system: a repressible binary...

  13. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... three components: a GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein...protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat protein...64543or 64542) When the reporter mRNA is first transcribed, it is bound by both the GFP and the RFP proteins...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ...pyralis) reporter plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...

  14. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase.... More detailed descriptions of the lentiviral elements and the 2nd and 3rd Generation Lentiviral Systems...Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning and...and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 puro (Plasmid...together with expression of fluorescent or enzymatic reporters for cellular assays and animal imaging studies...Additional Resources on the Addgene blog Viral vector topic page Beginners guide to viral vectors Quick guide...

  15. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following...The vast majority of fly vectors are based on p-element, random or targeted, integration into the fly genome...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g... the protein available – you can always add an epitope tag or a fusion protein. Mammalian vectors One ... allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently...e.g. pBAD LIC cloning vector (8A)) and have an epitope tag (6xHis or GST) to affinity purify the expressed...expression pattern of a defined promoter region, ectopically expressing YGOI in a pattern that allows the ...

  16. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...GCaMP6 family of sensors are commonly used calcium reporters with different kinetic parameters. There are three...ones? Many of the available biosensors use a single GFP or CFP/YFP-derived FRET pair. If you need to free...also available. Detection methods Many calcium reporters use a single fluorescent protein (FP), requiring...unwanted variables. Twitch is a FRET-based calcium reporter derived from troponin C. It is engineered with...for in vivo studies) There are several calcium reporters specially designed for the aims and requirements...cell body or the axon (GCaMP6 only). The genetic elements required for Drosophila studies have been added...Spring Harbor Protocols. 2012 Jun 1;2012(6):pdb-top069609. Palmer AE, Tsien RY. Measuring calcium signaling...

  17. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage...of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in the pool...activity of Cas13a becomes activated and the RNA reporter will be cleaved resulting in activation of the... the Cas13a molecule brings a much anticipated element of diversity to the CRISPR toolbox and will continue...the Addgene Blog Check Out Our CRISPR Featured Topic Page Learn about Anti-CRISPRs Multiplex Genome Editing...

  18. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many of which are available..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...-ome using a quick, affordable, and ubiquitous reporter assay. In their recent publication, Kroeze et ...and test constructs composed of complex genetic elements. To demonstrate the usefulness of GAMP, the Jacks...allows for easy and rapid targeting of complex elements to specific genomic sites. For instance, the authors... of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The...endogenous proteins. As described in their recent Cell Reports paper, the Kanemaki lab overcame this hurdle using...

  19. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...high is based on the reprogramming efficiencies reported in González et al. A percentage is not listed ...transposon: The PB transposon is a mobile genetic element that transposes via a “cut and paste” mechanism...plasmids carry the origin of replication (oriP) element and the cis-acting EBNA-1 from Epstein-Barr virus... iPSCs Find gene-specific plasmids (including reporters and RNAi) for:  NANOG  OCT4 SOX2 MYC KLF4 LIN28...generation of iPSCs is relatively simple in concept: ectopically express a cocktail of stem cell reprogramming...wide range of cell types and replicates in the cytoplasm independent of the cell cycle. A challenge of ...

  20. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ... (Dlx5/6) enhancer elements and combined it with a variety of reporters (i.e. eGFP) and effectors (i.e...phosphatase-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked mCherry...are available at Addgene. Kim, et al. Stem Cell Reports. 2015. PubMed PMID: 25772473 Targeting and manipulating... developed a new AAV approach using regulatory elements that restrict expression to this cell type. To...reagents shared via Addgene. Narayan S, et al. Cell Reports. 2017. PubMed PMID: 28813671 Listen to the podcast...along with EGFP while retaining an emission spectrum that is easily separable from that of EGFP. This makes...spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that...
Showing: 1 - 20 of 22 results