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We narrowed to 148 results for: GFP reporter gene

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  1. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...among other things) gene delivery, generating pluripotent stem cells, and developing gene therapies for monogenic...and lentiviral vectors that are used for gene regulation, gene of interest expression, knockout, and screening...gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for...for cloning and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...Experimental uses include: Cre-dependent gene expression, gene knockout, and selection marker removal. ...Expressing your gene of interest pLenti-puro (Plasmid #39481) You can clone your gene of interest into...

  2. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    ...epitopes, such as GFP (Paix et al., 2014). A further development of this approach reported in 2017 uses the... Mello lab also reported that modifying the 5’ modification of oligos used to generate linear dsDNA repair...where his group focuses on gene regulation in nematode molting and spermatogenesis. His group also builds ... one of the model organisms which I use to study gene regulation during developmental processes, CRISPR...drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes...Craig Mello’s lab – used inactivation of the unc-22 gene as their selection marker (Kim et al., 2014). Meanwhile...approaches may become even more powerful following a report that one can use linear repair templates (ie. PCR-derived...

  3. Hot Plasmids - August 2020

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    ...stem cells By Jennifer Tsang CRISPR gene interference (CRISPRi) and gene activation (CRISPRa) are great ways...GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato...into the hPSC genome using AAV1 and contain an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9...the plasmids at Addgene! A new tool for synthetic biology uses CRISPRi to control gene expression. This...nucleus and modulate transcription of the mCitrine gene. mCherry is constitutively expressed. Cells are ...selectivity and found that the best motif was from the Lck gene. For marking microtubules of mammalian cells, the... ways to modulate the expression of nearly any gene via specific guide RNA (gRNA) sequences. But these...

  4. 10 Great Guest Posts We're Thankful For

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    ...viral replication and transcription as well as reporter genes. To learn how you can use minigenomes in your...repression of your target gene, activation and repression of different genes in a single cell, and localization...Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts ...Lab at MIT developed plasmid based systems that report the order in which specific chemical inputs are...recombinase-based state machines could potentially be used to report upon and/or engineer “temporally ordered biochemical...derived from Zika virus and, upon binding, produce reporter proteins. These toehold switches can be used to... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (...

  5. Bikard Lab - CRISPR Repression Collection

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    ...levels for these two reporters simultaneously. By fusing sfGFP and/or mCherry to genes of interest, it is...expression of two genes without cross‐talk. (A) Schematic of the strain expressing two reporters and dCas9 under...attachment sites. The levels of the two reporters, mCherry and sfgfp in this case, can be controlled using...to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression of the RNA polymerase...enables robust, noiseless knockdown of bacterial genes. Molecular Systems Biology, 14(3):e7899. doi: 10.15252... the tracrRNA sequence (not shown). (B) Relative GFP and RFP concentration given relatively to the non‐targeting...carrying two CRISPR guides. The first one binds to sfgfp with either 0, 10, 11, 14, or 20 matching nucleotides...

  6. Fluorescent Proteins 101: Aptamer Fluorophores

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    Blog Post
    ...nature of GFP fluorescence to create an improved fluorescent aptamer system. Knowing that GFP’s  structure...Protein Activity with GFP Resources at Addgene.org Find the Spinach Plasmids at Addgene Find Broccoli Plasmids...Resources on the Addgene Blog Tips for Using FRET in Your Experiments Learn more about GFP Controlling Protein...highly customizable in terms of how they actually “report” as biosensors. Making aptamers glow The flexibility...increase its fluorescence, essentially mimicking GFP.  Though the aptamer, called Spinach, required longer...and used in assays for over half a century; the Addgene blog has an excellent primer and tips from experts...can be created in many different ways, but the general premise is that an aptamer in its unbound state...

  7. New Viral Vectors - Fall 2024

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    ...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...services collection or check out Addgene's Data Hub to see viral vector reports from other users. And don't...AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also have eight new AAV-PHP.eB...preps available in the Addgene viral vector repository! This includes the first Addgene prep packaged in the... AAV1 Optogenetics Karl Deisseroth New serotype pAAV-hSyn-hChR2(H134R)-EYFP AAV2 Optogenetics Karl Deisseroth... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...other new AAV vectors as well — including next-generation biosensors for GABA, dopamine, serotonin, and...

  8. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    ...irreversible PA-FP reported was PA-GFP. PA-GFP was derived from Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating...mutation in PA-GFP generates a chromophore population that is more neutral, causing PA-GFP to display almost...example, by engineering PA-GFP tagged to a mitochondrial matrix protein (mito-PAGFP) the Youle Lab was able...Multi-Color Imaging Catch Up on Your GFP Background Resources at Addgene.org Find Photoactivatable FPs in ...the neutral to the anionic form, which allows PA-GFP to exhibit up to a 100-fold increase in fluorescence...when illuminated with blue light (1). Similar to PA-GFP, scientists later developed red fluorescent PA-FPs...in vivo imaging models perfer the PA-RFPs over PA-GFP. Another class of irreversible PA-FPs are the photoconvertible...

  9. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether... promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...rescue genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing the...to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI’s... activator, bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including... to reduce the likelihood of indiscriminate reporter gene activation. Including a prey-only control also...

  10. Viral Production

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    ...vitro and in vivo for gene expression and/or function. These data are sometimes reported or posted on the ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, or both...Karl Deisseroth (Addgene viral prep # 55632-AAVrg). pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from... plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral helper sequences...recipient's initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...copies of the lentiviral Rev responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit...

  11. Another Pathway into Cells: iTOP

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    Blog Post
    ...targeting a gene associated with diphtheria toxin lethality and were sorted into single wells to generate clonal...PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters Previous Protein Delivery ...instance, it’s much more difficult to introduce a gene of interest into primary cells, so most primary ... 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,...Neuronal stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. ... risk of viral integration causing mutations and gene expression changes elsewhere in the genome. Second...

  12. Negative Can Be Positive: Open AAV Data with Addgene

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    ...Adeno-Associated Viral Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https...conditions commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https...neurons and GFP (green) to label neurons transduced by each serotype. Working at Addgene has shown ...It is a curated set of technical reports on AAV vectors from Addgene and the CLOVER Center, each one submitted...submitted by researchers like you. Reports are reviewed by Addgene scientists for clarity and accuracy... cassette with an ubiquitous promoter to express GFP in a group of young rats. We found that, in this ...Grames, M. S., & Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector dose–...

  13. Choosing Your Fluorescent Proteins for Multi-Color Imaging

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    ...imaging with this set are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T, and an...with this filter set and have reported brightness measurements. Here, EGFP outperforms the improved folding... / em 527 nm) can be imaged together but CFP and GFP (ex 488 nm / em 507 nm) show some crosstalk between...TagRFP to give a brighter signal than mCherry. In general, filter sets designed for Rhodamine / Cy3 will ...mammalian cells, one of the improved folding variants of EGFP like mEmerald or Clover is probably best; mNeonGreen...set of proteins the green and red proteins are generally the most detectable and so should be used to tag...

  14. Lentivirus Plasmids

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    ...promoter for transgene and CMV drives expression of RFP as a reporter. See plasmid 17618 for GFP plasmid. ...viral particle) and toxicity of the gene product (i.e., if the gene encoded by the transfer plasmid is ...hUbC-driven EGFP; can be used for cDNA expression Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and shRNA ...Expresses shRNA under mouse U6 promoter; CMV-EGFP reporter cassette is included to monitor expression Parijs...Expresses shRNA under mouse U6 promoter; CMV-EGFP reporter cassette is included to monitor expression Kissler...24129 pULTRA 3rd bi-cistronic expression of EGFP and the gene of interest, also can express shRNA (cloning... which can be modified to encode a wide range of gene products. The main restrictions on a lentiviral ...

  15. Plasmids 101: Mammalian Vectors

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    Blog Post
    ...lines and are not used for general plasmid selection. Instead, reporter genes are often employed to easily...test for your gene of interest may be another method to assess transfection success. GFP is often used...expression levels in the cells. Ideally, the chosen reporter is unique to the cell, is expressed from the plasmid...used as a reporter and we will be covering this ever-versatile fluorophore in a later post, so stay tuned...conjunction with positive selection to ensure your gene gets targeted to a specific location within the ...used selection markers are listed below: Name Gene Conferring Resistance Cell Types* Mode of Action...use G418. This can be confusing since the neo/kan gene confers G418 resistance; however, like gentamicin...

  16. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

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    Blog Post
    ...fluorescent protein reporter under the control of promoters that control cell cycle gene expression.  Additional...Resources on the Addgene Blog Visit the fluorescent protein topics page Get the basics on GFP Find microbiology..., Bernach plans to use a suite of fluorescent reporters. Bacteria containing unstable fluorescent proteins...colonizer, labelled with a constitutively fluorescent reporter can also be used to track how one population responds...work in Lin Tian’s lab at UC Davis is to generate a genetically encoded fluorescent sensor for serotonin... the TIMER growth rate bioreporter. During protein maturation, the bioreporter spontaneously changes from...sequence an individual’s genome. But when they find genetic variants, what does that mean? Previous research...

  17. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ...both introduce genes into an organism’s genome (see figure) and to disrupt endogenous genes at the site ...puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each contain... fused to a region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new...first-generation transposase (as determined by transposition assays of antibiotic-resistance genes conducted...). In their Nature Genetics paper, the lab showed that efficiencies of stable gene transfer using SB100x... the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest... exciting tool for producing specific genetic mutations and gene disruptions without the use of viral ...

  18. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    ...and a mCherry reporter downstream of the PBAD promoter. To use this plasmid with your gene of interest,...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...interest, you would replace mCherry with your gene of interest. pBLADEONLY: This plasmid contains only BLADE ...already have a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform ...co-transform pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ... Resources on Addgene.org Check out the Molecular Biology Reference Download Addgene's Plasmids 101 eBook...

  19. Hot Plasmids: Summer 2024

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    ...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...Prime-editing-Assisted Site-Specific Integrase Gene Editing (PASSIGE), their previously-reported technique for precisely integrating...editing, this epigenetic strategy doesn’t induce DNA damage or permanently alter a gene product; it simply...In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are captured...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines ...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...

  20. Choosing Your Perfect Empty Backbone

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    ...popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). ...These vectors allow you to test the function of Your Gene Of Interest (YGOI) in a controlled environment under...design. You can create a new strain with a disrupted gene by transforming wild-type yeast with an integrating...visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected cells... More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other...

  21. Viral Vectors 101: Calcium Sensors

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    Blog Post
    ...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...Resources on Addgene.org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’s Viral Services...calcium sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent...

  22. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Blog Post
    ...of where the genes were expressed. What’s the use of stable gene expression? Constant gene expression can...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells...input and gene expression is the output. The input, plasmid copy number, directly influences gene expression...buffered against the effects of copy number on reporter gene expression. On the other hand, using a constitutive... varied in size or carried different genes. In these cases, gene expression also remained stable regardless...media and growth stage can both affect gene copy number. Moreover, gene copy number is affected differently... constitutive promoter upstream of GFP increased expression of GFP six-fold, but a stabilized promoter...

  23. Hot Plasmids - February 2022

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    ...to induce high levels of gene activation for both reporter and endogenous genes. It is highly specific ...anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling...between MARCH5 and WSB2 genes using Cas12a’s ability to knock out multiple genes in parallel. Lastly, this...cellular targets Optimized genetic screening technique using Cas12a for multiplexed gene perturbation The Penn...10846-1.   Optimized genetic screening technique using Cas12a for multiplexed gene perturbation by: Gabrielle...many CRISPR-based genetic screening methods have primarily used Cas9 to knock out genes, researchers at ...multiple genes within a single cassette provides increased simplicity for combinatorial genetic screens...

  24. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

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    ...limits the delivery of larger transgenes, such as base editors which have a gene size of ~5.2 kb. To overcome...Ace-mScarlet voltage indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo...Article contributed by Jennifer Tsang AAVs are a great gene therapy delivery mechanism, but their small packaging...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...dosage comparable to those currently used in human gene therapy trials. They further tested the base editors...voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...expression levels from different promoters, and inhibited gene expression using CRISPRi. The kit functions in the...

  25. Technologies Enabled by NanoLuc® Luciferase

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    ...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged... bright enough to detect endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc...imaging tools generate cyan, yellow, green, orange and red fluorescence. The paper reports a wide variety...photoactivatable, optogenetic accuators to depolarize or hyperpolarize membranes. Traditional FRET reporters could...would also activate the optogenetic accuators. The researchers took a FRET reporter and adapted it into a...Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are available...couples NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...

  26. Lighting Up Cell Signaling with Photoswitchable Kinases

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    ...Control Protein Activity with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable... an ERK kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR...spatiotemporal resolution than pharmacological or genetic approaches, but only a few such methods exist and...Solution: In Zhou et al, the Lin lab present a generic design for creating photoswitchable kinases and...possible to create other photoswitchable kinases? The general design of psMEK1 also worked for creating three...PMCID: PMC4097317. Additional Resources on the Addgene Blog Learn More about Photoactivatable Fluorescent...

  27. Serotype Testing AAV

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    ...Serotype Testing AAV These AAV encode fluorescent reporters and can be used to compare the tropism of different...AAV1). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in...plasmid 37825 (deposited by Edward Boyden ) and direct GFP expression from the CAG promoter. For information...Ready-to-use AAV available from Addgene's viral service. Control EGFP vectors in various serotypes for... 37825-AAVrg.T 20 µL $ 150 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV ...plasmid 50465 (deposited by Bryan Roth ) and direct EGFP expression from the human synpasin promoter. For...

  28. Lentiviral Prep Service

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    Collection
    ...Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ...on each item's material page. Actual titers are reported with each shipment. For more details about our...Screening Use pooled CRISPR libraries to screen for genes involved in specific biological processes. For more... Plasmid Pooled Libraries . ID Name Description Genes/Insert Selection PI Human knockout pooled libraries...library in backbone lentiGuide-Puro targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000...SpCas9. 76,441 unique sgRNAs targeting 19,114 human genes along with 1000 non-targeting controls Puromycin...

  29. pSiM24: Simplifying Plant Genetic Engineering

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    ...and corn. For both β-glucuronidase (GUS) and GFP reporters, as well as multiple transient and transgenic...ideal for large gene transfer. The M24 promoter provides constitutive, high-level gene expression and ...further customizing gene expression. Sahoo et al. have tested the system with genes up to 4 kb in length...versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants...Scientists have worked to optimize this system for gene transfer, studying the stability of modified Ti plasmids...versatile binary Ti vector for both transient and stable gene expression applications in plants. This smaller,...and many plant scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity...

  30. Brain Initiative Collection

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    ...Fishell 83895-AAV1 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV2 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV8 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV9 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...element Gordon Fishell 83900-AAV1 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV2 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...

  31. Simplify Cloning with in vivo Assembly

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    ..., or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications...insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer design for each type...pathway is ubiquitous, with successful recombination reported in all laboratory E. coli strains tested so far...use of this pathway as a cloning tool was first reported over 25 years ago (Douglas and Howard, 1991, Bubeck...amplified, restriction digested or even a synthesized gene. PCR is the primary method to produce the linear...included in a single PCR, for example for subcloning genes from different templates. All PCRs are run as an... 5’ end of either primer. Subcloning - To move a gene from one vector to another simply amplify the vector...

  32. Cre-Lox and Other Site-Specific Recombinases

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    ...excised, and gene expression proceeds. Gene Knockout: Placing the loxP sites on either side of a gene (called...shRNA expression. Gene Switch: These constructs contain two genes of interest, Genes A and B. When Cre... few popular recombinase-dependent gene expression strategies. Gene Expression: A stop codon flanked with...LSL" cassette) placed upstream of a gene of interest will prevent gene expression in the absence of Cre.... loxP"), will permit gene expression until Cre is present, at which time the gene will be disrupted or...Cre is absent, only Gene A is translated correctly. Cre expression excises Gene A and alters the reading...backbone to regulate your gene of interest. Depending on the construct, gene expression may be activated...

  33. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    ... developed a third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study...research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research ...line, Hep3B. From left to right: Mitotracker Red, GFP for the biosensor, Hoechst, and overlay).   ...(DKFZ) who has developed new redox sensitive GFP2 (roGFP2)-based biosensors for live cells. His work focuses..., Pedre Pérez’s biosensor fuses MPST to roGFP2. Because roGFP2 contains two cysteines near the chromophore...developed a dual virus system using both optogenetics and chemogenetics to study the activity of parvalbumin...catalyzes the desulfuration of 3-mercaptopyruvate and generates an enzyme-bound hydropersulfide that is then transferred...

  34. Chemogenetics AAV Preps

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    ... 52536 rAAV-CAG::FLEX-rev:: hM4D-2a-GFP hM4D(Gi) - Inhibition GFP Cre-dependent 1 Sternson 154867 pAAV-hSyn-fDIO-hM4D...PHPeB Clear Filters ID Name Receptor and Activity Reporter/Tag Cre requirement Serotype PI 44361 pAAV-hSyn-DIO-hM3D...Service AAV Chemogenetics Viral Vector Packaging Service: Chemogenetics AAV Use our chemogenetics AAV to chemically...IRES EGFP PSAM4 GlyR - Inhibition IRES EGFP none 5 Sternson 119744 AAV CAMKII PSAM4 GlyR IRES EGFP PSAM4...Ready-to-use AAV available from Addgene's viral service encoding chemogenetics tools. Viral...in specific cell types. See our Chemogenetics Plasmids and Chemogenetics Guide pages for more information...119741 AAV SYN flex PSAM4 GlyR IRES EGFP PSAM4 GlyR - Inhibition IRES EGFP Cre-dependent 5, 9 Sternson 119742...

  35. Plasmids 101: What is a plasmid?

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    Blog Post
    ...typically include a gene that is advantageous for survival, such as an antibiotic resistance gene. In contrast...the target gene. Vital component for expression vectors: determines which cell types the gene is expressed...cloning plasmids, expression plasmids, gene knock-down plasmids, reporter plasmids, viral plasmids, and genome...scientists use plasmids? Generally, scientists use plasmids to manipulate gene expression in target cells...replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing bacteria...MCS is often downstream from a promoter. Insert Gene, promoter or other DNA fragment cloned into the ...obtained. Selectable Marker The antibiotic resistance gene allows for selection in bacteria. However, many ...

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

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    Blog Post
    ...include a reporter like GFP, some smaller CRISPR-Cas proteins such as SaCas9, CRISPR gRNAs, or your gene of ...through this challenging sequence. Transgene The transgene is the gene that will be delivered by the AAV...expression of the AAV transgene. On a plasmid map, promoters are upstream or 5’ of the gene they control. Promoters...plasmid which contains the structural and packaging genes, the adenoviral helper plasmid which contains the...for the successful and specific expression of your gene of interest. References Haery L, Deverman BE, ...Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods 30:206–213 . https://doi.org/10.1089...control transgene expression. Lox sites are directional 34 bp sequences that can flank the transgene and ...

  37. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    ...following sucrose treatment. Moreover, several reports have shown that cells treated with sucrose or other...endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...during adeno-associated virus manufacturing." Human gene therapy21.10 (2010): 1251-1257. PMID: 20649475. ...have probably heard about Addgene’s viral service but did you know that Addgene also performs independent...101 eBook Additional resources on Addgene.org Learn about Addgene's viral service Learn about viral production...research? Since launching the viral service in 2016 Addgene’s research team has been actively investigating ...PMCID: PMC2957237. Additional resources of the Addgene blog Read our beginner's guide to viral vectors...

  38. Caltech Systemic Capsids

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    ...Dimidschstein 135631 pAAV-S5E2-GFP-fGFP E2 regulatory element GFP, membrane-targeted GFP Control Dimidschstein ...pAAV-CAG-GFP CAG GFP Control Boyden 50465 pAAV-hSyn-EGFP hSyn EGFP Control Roth 50469 pAAV-CaMKIIa-EGFP CamKIIa...104061 CAG-NLS-GFP CAG NLS-GFP Control Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Control Wilson...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden MaCPNS2 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden CAP-B10 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden CAP-B22 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden AAV9-X1.1 These viral vector preparations...

  39. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted GFP into an intracellular loop...of interest and co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma...And orthogonal sensors with different ITAM/SH2 reporter pairs allow multiplexed measurement of different...10.1101/2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent proteins promise...photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and its small size (~1500...new to Addgene this year. Feel the love with MTRIA sensors One of the latest additions to Addgene’s catalog...imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator using small...

  40. New Viral Vectors - Winter 2025

    Type
    Blog Post
    ...multiple) rAAV-CAG::FLEX-rev:: hM4D-2a-GFP AAV1 Chemogenetics Scott Sternson New viral prep pAAV-CaMKIIa-hM3D...to request? Addgene's Data Hub can help, with over 100 user-submitted viral vector reports. And don't ...mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-CaMKIIa-hM4D(Gi)-mCherry AAV1 Chemogenetics Bryan ...mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-hSyn-hM4D(Gi)-mCherry AAV1 Chemogenetics Bryan Roth...AAV-PHPeB Optogenetics Jonathan Ting New viral prep AiP14033: pAAV-AiE0779m_3xC2-minBG-CoChR-EGFP-WPRE3-...AAV-PHPeB Optogenetics Jonathan Ting New viral prep AiP14035: pAAV-AiE0452h_3xC2-minBG-CoChR-EGFP-WPRE3-...pAAV-AiE0873m_3xC2-minBG-CoChR-EGFP-WPRE3-BGHpA (Alias: CN5140) AAV-PHPeB Optogenetics Jonathan Ting New viral...

  41. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...Plasmid 68295 GFP E17TAG Plasmid 68296 GFP S2TAG Plasmid 68297 GFP Q157TAG Plasmid 68298 GFP S2TAG/E17TAG...useful for the incorporation phosphoserine into your gene of interest. Depositor...expression vector encoding a split mCherry fluorescent reporter, enabling the detection of phosphorylation-dependent...E17TAG Plasmid 68299 GFP E17TAG/Q157TAG Plasmid 53225 MBP-MEK1 S218TAG/S222TAG ( aka. PCRT7 tetR pLtetO MBP-MEK1...68305 Beta lactamase S68TAG Plasmid 69118 E17TAG GFP zeo resistance Strain 34929 BL21ΔserB Strain 52055...by adding this phosphorylated amino acid to the genetic code of E. coli (Park et al., Science 2011). Follow... in Mohler et al., Mol Syst Biol 2023 and find general tips on phosphoprotein expression in rEcoli XpS...

  42. Hot Plasmids February 2024

    Type
    Blog Post
    ...Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter (mChe)...-invasive systemic gene delivery in non-human primate (NHP) brains.  Systemic gene delivery capsids previously...Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific .... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain....plasmid here! Chen, X., et al. (2023). Functional gene delivery to and across brain vasculature of systemic...past decade, primary cells remain difficult to genetically edit due to toxicity from electroporation or ...

  43. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...expression of reporter genes, or to combine reporter expression from one promoter with transgene or siRNA ... transcription of a target gene, it therefore determines the timing of gene expression and largely defines...These permit exquisite control of gene expression and tracing of gene expression across development. One...stage-specific promoter and design a UAS reporter construct, the reporter will only be expressed where and/or...of herpes simplex virus immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101...systems are genetic tools that consist of two parts, both of which must be present for a gene of interest...placing UAS upstream of a transgene permits directed expression of that gene in cells that also express...

  44. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...Resources on the Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably...expression time. 2. Timer functionality with a single gene Although it is possible to make a timer system composed...cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...FP timer DsRed-E5 under the control of Neurog3 (a gene controlling pancreatic differentiation) in mouse...(trafficking to a subcellular location, start of gene expression, development of a cell structure, etc...formation of limbs are correlated with changes in gene expression. In a developing embryo, for example,...

  45. Genetic Code Expansion

    Type
    Collection
    ...express first with a control reporter gene – GFP for E. coli or mCherry-GFP for mammalian cells. You should...EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR). ...EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tolC, and tyrU gene replaced by GentR) and...Removed endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV gene replaced by tRNA variant that suppress...endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV and tyrU genes replaced by tRNA variant that suppress...endogenous EcTyrRS/tRNA pair (tyrS gene knocked out, tyrTV and tyrU genes replaced by tRNA variant inactivated...that contain genes with modified codons for unnatural amino acid incorporation. ID Plasmid Gene/Insert Vector...

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

    Type
    Blog Post
    ... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage... (Shmakov et al., 2015). They were using Cas1, a gene commonly associated with CRISPR arrays and involved...analysis, they identified 53 potential candidate genes that fell into 3 categories based on the architecture...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...expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although the...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check Out Our CRISPR Featured...

  47. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...destination vector contains GFP, enabling you to select cells with high GFP expression. These cells have...sequence between two target sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple...plasmids, which contain various promoters and reporters, and subsequently inserted into a Cas9-containing...simultaneous multiplex editing of six different genes. This system doesn’t rely on any additional or host-dependent...-in-one CRISPR/Cas9 vector system.” Scientific Reports 4 (2014): 5400. PubMed PMID: 24954249. PubMed Central...from this paper at Addgene. 9. Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic ...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Browse Our CRISPR Topic...

  48. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red light rapidly deactivates...plasmids from this publication at Addgene. Multichromatic control of gene expression in Escherichia coli...optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...used to engineer new pathways with sophisticated gene expression controls. Most cell types do not respond... can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive two-component... was only about 10-fold, precluding the study of gene expression at very high or very low levels. Schmidl...researchers to better fine-tune the level of output gene expression based on the input light intensity. In...

  49. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...µg/mL Anti-GFP [N86/38.1R] (Addgene #180084), 0.01 µg/mL Anti-HA [12CA5] - Chimeric (Addgene #198003)) ...express either the optogenetic activator channelrhodopsin-2 or fluorescent reporter dTomato under the ... intensity-based Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren ... Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible tagging:...tag. Selectable and stable: A puromycin resistance gene makes stable cell line creation a breeze. Modular...labeling By Alyssa Shepard To fully understand how genes are regulated, it's important to know what proteins...identify both new and known regulators at the FOS gene promoter and the MYC locus (Figure 5). Figure...

  50. Neurodegeneration Research Collection

    Type
    Collection
    ...type or mutant ORFs, gene fragments, and gRNAs for genes linked to neurodegenerative disorders. Find plasmids...within the huntingtin (HTT) gene. An individual with the mutated HTT gene will have huntingtin proteins...association between a gene and late-onset AD. It is more likely a combination of gene variation and lifestyle... are associated with known genetic mutations in one of the following genes: SOD1, C9ORF72, TARDBP, FUS...three different inherited genes: Presenilin 1, Presenilin 2, and APP gene.The majority (>90%) of individuals...these additional genes may play in Alzheimer's disease. Research Tools Neurodegeneration Plasmid Collection...FRET-based reporter to study tau seeding in mammalian cells. Lathuiliere et al. Mol Neurodegener. 2023 Aug...
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