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  1. Choosing Your Fluorescent Proteins for Multi-Color Imaging

    Type
    Blog Post
    ...best fluorescent proteins for imaging with this set are mTagBFP2, EGFP or one of the improved GFP variants...follow multiple fluorescently tagged species simultaneously. To do so with fluorescent protein labels ...in distinct fluorescent channels on the microscope. With the proliferation of fluorescent proteins in ...choice of fluorescent proteins requires some thought. The first step in choosing fluorescent proteins...is to be aware of what fluorescent proteins are available. With new fluorescent proteins being published...latest fluorescent proteins, I maintain an interactive graph and table of the best fluorescent proteins...available. Choosing compatible fluorescent proteins To choose a set of fluorescent proteins to be imaged together...
  2. Hot Plasmids and Viral Preps - September 2021

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    Blog Post
    ...mCherry fluorescent protein followed by a stop codon and the coding sequence for a green fluorescent protein...representation of cell-free system, capable of producing fluorescent proteins and enzymes. Image from Moore et al....validated through monitoring the expression of fluorescent proteins and oxytetracycline enzymes. Next, this...plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE, to identify...protein (GFP). The success of the A-to-G conversion could be seen through the success of the change in expression... expression of GFP, for the stop codon should have changed from “TGA” to “TGG.”  After identification ...cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s. b) Quantification ...
  3. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    ... to use a suite of fluorescent reporters. Bacteria containing unstable fluorescent proteins under the ...to reduced eGFP fluorescence. Cells expressing the library are then sorted by fluorescence-activated cell...inoculation, no more fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be...colonizer, labelled with a constitutively fluorescent reporter can also be used to track how one population...growth rate using TIMER and an unstable fluorescent protein reporter under the control of promoters that ...Addgene Blog Visit the fluorescent protein topics page Get the basics on GFP Find microbiology blog posts...wild-type levels of abundance, the eGFP will be stable and fluoresce. But if the variant is unstable, it...
  4. 10 Great Guest Posts We're Thankful For

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    Blog Post
    ...Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to ...Janelia Research Campus reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores...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...viral replication and transcription as well as reporter genes. To learn how you can use minigenomes in...to different pH’s and some that are fused to fluorescent proteins. She also showed that R-bodies can rupture...
  5. Cre-lox system

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    Collection
    ...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG...116879 CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral...expressed in the retina). Fluorescent Cre: The fusion of Cre to a fluorescent reporter enables visualization...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...13770 pCALNL-GFP Cre dependent GFP expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP-dsRed gene switch...promoterless CRE-GFP fusion none Mammalian Sauer 11960 pBS537 tet-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV...Nestler 68552 HRE:GFP-pGK:Cre Cre and hypoxia-responsive GFP reporter HRE Lentiviral Jacks 69570 pAAV-EF1a-...
  6. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...better biosensors. NanoLuc®-fluorescent protein fusions for imaging Fluorescent proteins are fantastic imaging...more penetrative orange fluorescent protein fusion (OgNLuc). Both fluorescent proteins can be excited ...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged...for infection of host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected...set out to design a better fluorescent protein that could be co-imaged with GFP using the same wavelength...cyan, yellow, green, orange and red fluorescence. The paper reports a wide variety of applications of the...
  7. Viral Vectors 101: Calcium Sensors

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    Blog Post
    ...measurements, specifically fluorescent ones? Many of the available biosensors use a single GFP or CFP/YFP-derived...available. Detection methods Many calcium reporters use a single fluorescent protein (FP), requiring only a simple.... This sensor switches from green fluorescence to a red fluorescent state only by simultaneous (1) user-controlled...Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (... (e.g., calmodulin or troponin C). Their fluorescent readouts are used to monitor calcium concentrations...GCaMP6 family of sensors are commonly used calcium reporters with different kinetic parameters. There are three... of the sensor, and (3) subsequent change in fluorescence only occurs at high calcium concentrations. ...
  8. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    Blog Post
    ... microbes rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue ... consists of a library of reporter plasmids, each containing two fluorescent proteins and four repeats...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...and a reporter for tracing the engineered cells in vivo, the non-immunogenic violet-excited GFP variant...Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists...Mikhail Shapiro’s lab has developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression...
  9. Fluorescent Proteins 101: Fluorescent Protein Timers

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    Blog Post
    ...time.Hallmarks of fluorescent protein timers When FP timers are first expressed, they predominantly fluoresce one ...independently) originally fluoresce green or blue and, when they mature, fluoresce red. The ratio of the ...., et al. "Monomeric fluorescent timers that change color from blue to red report on cellular trafficking...Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing...Choosing Your Fluorescent Protein Resources at Addgene.org Find Fluorescent Protein Timers Browse All...All Fluorescent Protein Empty Backbones Check Out Our Fluorescent Protein Guide ...Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell...
  10. Plasmids 101: Control Plasmids

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    Blog Post
    ...plasmid that constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected...transfection was not successful due to the absence of GFP fluorescence from the living, viable cells. This result... well of your cells. Regardless, the amount of reporter protein activity correlates to both the amount...result in Figure 1, an internal control, such as the GFP-expressing Plasmid B, could demonstrate whether the...successfully and expressing the protein. For example, fluorescence microscopy images resulting from our experiment...produces the expected effect, which is green fluorescent cells (Figure 3).   Figure 3: Expression of Plasmid...
  11. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...when paired with appropriate fluorescent optics. Existing red fluorescent voltage indicators, however,...levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo and achieved...31937940. Detection of neural activity using a red fluorescent genetically encoded voltage indicator Article...however, fall short compared to GFP-based sensors in terms of response and brightness. They also do not spectrally...voltage-sensitive inhibitory rhodopsin (Ace2N) with the red fluorescent protein (mScarlet) with either a seven or eight...manipulations when combined with existing green-fluorescent GEVIs and blue-light-sensitive channelrhodopsins...indicator plasmids at Addgene! Beck et al., Sci Report 2019. PubMed PMID: 31685893. A MoClo toolkit for...
  12. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ...pPS808 - N-terminal GFP for yeast expression pET GFP - C-terminal GFP for...gene expression GFP Varies pBABE GFP - Retroviral gene expression pMKO.1 GFP - Retroviral ...C elegans expression GFP Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression... from the SP6 promoter pCS2 mt-GFP - Create myc-tag, GFP-tag fusions for transcription ...expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian expression...pPD95_75 - C-terminal GFP for C elegans expression Other Fluorescent Proteins Localization...expression MSCV-IRES-GFP - Mammalian retroviral gene expression with GFP selectable ...
  13. Lighting Up Cell Signaling with Photoswitchable Kinases

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    Blog Post
    ... an ERK kinase translocation reporter with a mRuby2 fluorescent reporter (ERK KTR-mRuby2). The ERK KTR...with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins... the active site. Dronpa also produces green fluorescence when exposed to 500 nm light.  Under 400 nm ...active, it phosphorylates the ERK KTR and red fluorescence localizes to the cytoplasm. When MEK1 and ERK...inactive, the ERK KTR isn’t phosphorylated and red fluorescence can be found in the nucleus. For testing kinase...Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox...Photoswitchable Fluorescent Proteins ...
  14. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    Blog Post
    ...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used... to study gene expression in cells. But these reporters have limited use in vivo because they are based...expression in vivo.  But what ultrasound lacked was reporter genes that could produce ultrasound contrast. ...our lab - the Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when...without signs of toxicity or burden relative to a fluorescent protein control. We also added the toxin-antitoxin...P2A-linked accessory genes gvpNJKFGWV marked with Emerald GFP. These plasmids can be used directly in transient...gating for the top quartile (or higher) of BFP and GFP producing cells. After FACS, return the cells to ...
  15. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ... blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent protein... co-expressing a cytosolic ZtSH2 reporter, they observe fluorescence localizes to the plasma membrane ...2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent proteins promise better performance...area with lots of bright new plasmids is our Fluorescent Biosensors Collection — scientists are always...measure! Let’s take a look at some of the hottest fluorescent biosensors new to Addgene this year. Feel the...Hiroshi Hibino and colleagues devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular...region-to-IntrAcellular loop linkers” (MTRIA for short), where a fluorescent protein is inserted into one of the loop regions...
  16. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    ...live cells are GFP+. In this plot, with adjusted voltage, cells that did not receive the GFP protein are ...populations or a huge smear across your FSH vs reporter plot. In this post, I’ll walk you through how ...-case result. For example, you have introduced a GFP protein into your cells (visualized via the FITC ... to be clustered at signals below 103, while the GFP+ population cluster would be shifted to the right...samples and controls were processed the same way, the GFP- population should overlap with the negative control...control population (Figure 1). Based on where the GFP- population ends, you can draw a gate which selects...selects anything to the right of it, marking it as GFP+. Quantification for such graphs could simply be the...
  17. Brain Initiative Collection

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    Collection
    ...pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 AAV production plasmid encoding for Archon1 fluorescent voltage reporter, Cre-dependent...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...
  18. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...third-generation intensiometric glutamate sensing fluorescent reporter (iGluSnFR3) to study glutamate, a signaling...can be transferred to the roGFP2 cysteine thiols and change its fluorescence properties. The biosensor... for awards to early career scientists using fluorescent proteins in their research. These awards help...roles in the development and dissemination of fluorescent protein tools. This year, we’ve made some changes...research.   Abhi Aggarwal: Glutamate sensing reporters Abhi Aggarwal is a postbaccalaureate research ...disease,” says Aggarwal. This biosensor changes fluorescence based on the release of glutamate from neurons...line, Hep3B. From left to right: Mitotracker Red, GFP for the biosensor, Hoechst, and overlay).   ...
  19. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ... transformation. Choose good reporter genes There are many great reporter genes available and which one...pSB166 - pL2_pSB90_tMAS::rLUC-I::pMAS. Fluorescent proteins (FP, e.g. GFP) can be used in many ways. Often ...Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression studies in plants...the T-DNA region with our gene of interest or a reporter gene so that the transformation and gene expression...different treatments on transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase...histochemical GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate...transformation success. Luciferases are great quantitative reporters and use a substrate to produce bioluminescence...
  20. Luciferase Plasmid Collection

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    Collection
    ...Multiple Multi-luciferase reporter vector including transcriptional reporters for NF-kb, TGF-b, c-Myc, ...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson...transcriptional regulation and bioluminescent reporters. Plasmid...Luciferase Plasmid Collection You may also like... Fluorescent Proteins Blog: Luciferase 101 Blog: Technologies...Highlights Empty Backbones Expression Constructs Reporter Constructs Luciferase is the enzyme responsible... common choice in assays ranging from use as a reporter gene in vitro and in vivo to use in biosensors...-lives, making them useful as transcriptional reporters. However, they are not secreted, so cells must...
  21. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...will not produce the fluorescent protein while areas that are exposed will fluoresce. With these images,...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...contains BLADE, the PBAD promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid...strain transformed with pBLADE expressing superfolder GFP instead of mCherry under the PBAD promoter. They ...
  22. Lentiviral Prep Service

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    Collection
    ...express fluorescent proteins. ID Name Insert Antibiotic Resistance Purpose PI 17446 pLenti CMV GFP Hygro... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 .... This library expresses EGFP for easy visualization via direct fluorescence. Morris 115644 Morris CellTag.... This library expresses EGFP for easy visualization via direct fluorescence. Morris 115645 Morris CellTag.... This library expresses EGFP for easy visualization via direct fluorescence. Morris Control Lentivirus...Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV promoter, Hygro Campeau Viral...on each item's material page. Actual titers are reported with each shipment. For more details about our...
  23. Viral Production at Addgene

    Type
    Blog Post
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...expression and/or function. These data are sometimes reported or posted on the material page for the corresponding...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ...prep # 55632-AAVrg). pAAV-CAG- FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...recipient’s initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...
  24. New Viral Vectors - Fall 2024

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    Blog Post
    ...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also have eight new AAV-PHP.eB...drive expression of either Cre recombinase or a fluorescent marker in the various cell types of the basal...check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the link...Controls Jonathan Ting New viral prep pAAV-CaMKIIa-EGFP AAV1 Controls Bryan Roth New serotype pAAV-Ef1a-DIO...
  25. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ...stability of fluorescence over time is crucial. For instance, the fluorescence of fluorescein isothiocyanate...the b(right)est fluorescent proteinFluorescent Proteins 101: A History of Fluorescent Proteins  ...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes,...dyes, and fluorescent ligands. Figure created with biorender.com. Fluorescence refers to the emission...selection and optimization. Fluorescent experiment types: Different types of fluorescent experiments, such as...to the ability to ‘toggle’ between fluorescent and non-fluorescent states in response to a specific stimuli...technologies in fluorescent dyes/ligands Throughout the history of cell biology, fluorescent labeling techniques...
  26. Hot Plasmids February 2024

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    Blog Post
    ...Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter (mChe)...marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons...colleagues identified a highly bright and stable fluorescent protein, StayGold. Although it quickly saw wide...dimerization while maintaining the bright and stable fluorescence of the original (Ivorra-Molla et al., 2023)....Ivorra-Molla, E., et al. (2023). A monomeric StayGold fluorescent protein. Nature Biotechnology, 1–4. https://doi.org..., et al. (2023). Bright and stable monomeric fluorescent proteins derived from StayGold. ResearchSquare...
  27. Multiple Plasmids at a Low Price: Kits from Addgene

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    Blog Post
    ...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013... Using an Orthogonal Ribosome-Controlled Fluorescent Reporter."ACS chemical biology 11.1 (2015): 31-37...various combinations of protein tags (His, Avi, and GFP) as well as protease sites to facilitate protein ...library by in vivo functional screen." Scientific reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted...’ end of the leader sequence. Using a variety of GFP and RFP fusions as their test genes, the researchers...fusions. Some applications of PUF protein fusions reported in the literature include RNA detection, RNA cleavage...et al 2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison...
  28. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
    ...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...Cre), together with expression of fluorescent or enzymatic reporters for cellular assays and animal imaging...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...
  29. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    Blog Post
    ...the widely used green fluorescent protein (GFP) gives a bright green fluorescence when exposed to blue ...like chromoproteins, fluorescent proteins, and luminescent proteins. While fluorescent proteins and luminescence...from fluorescent proteins? Before going into the difference between chromoproteins and fluorescent proteins...light, whereas chromoproteins do not fluoresce at all or fluoresce at very low levels. Chromoproteins in...The ocean is full of striking visuals from fluorescent jellyfish, bioluminescent phytoplankton, and colorful... chromoproteins are actually a subset of the fluorescent protein family. What sets them apart is the way... in nature Compared to the history of fluorescent proteins in molecular biology research, chromoproteins...
  30. Serotype Testing AAV

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    Collection
    ...: Serotype Testing AAV These AAV encode fluorescent reporters and can be used to compare the tropism of...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... available from Addgene's viral service. Control EGFP vectors in various serotypes for serotype testing... 37825-AAVrg.T 20 µL $ 140 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...
  31. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    Blog Post
    ... Posts on Fluorescent Proteins Browse Our Fluorescent Protein Pages Find the mOX Fluorescent Proteins ...protein non-fluorescent. Unfortunately, accumulated misfolded and nonfunctional (dark) fluorescent proteins...Costantini, and Erik L. Snapp. "Superfolder GFP is fluorescent in oxidizing environments when targeted via...applicable to many microscopy experiments with fluorescent proteins (FPs) localized to compartments other...that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions on quantitative imaging... and cell physiology in general. Pitfalls of Fluorescent Protein Fusions Observing a bright signal for... fail to correctly fold? Misfolded FPs do not fluoresce. A dark population is not readily apparent and...
  32. Viral Production

    Type
    Collection
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...expression and/or function. These data are sometimes reported or posted on the material page for the corresponding...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ... prep # 55632-AAVrg). pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...recipient's initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...
  33. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...neomycin, puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each...region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new tool kit allows...
  34. Choosing Your Perfect Empty Backbone

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    Blog Post
    ...popular studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following...in-frame fluorescent fusion protein will allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you...animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g...
  35. The Fluorescent Vegetables in Aptamer Soup

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    Blog Post
    ...small molecule activates the fluorescence of the small molecule. Fluorescent RNA aptamers are used to track... applications of fluorescent RNA aptamers in Addgene’s collection. Green fluorescent aptamers Spinach ...selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution...using aptamers might know this best, as most fluorescent aptamers are named for a fruit or vegetable! ...What is an RNA aptamer? RNA is not inherently fluorescent and can be challenging to track intracellularly... affinity to small molecules to induce their fluorescence. Until a binding event occurs, neither the aptamer... aptamer nor the small molecule are strongly fluorescent but the binding of an aptamer to its target small...
  36. Rinehart Lab Phosphoprotein Reagents

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    Collection
    ...Plasmid 68295 GFP E17TAG Plasmid 68296 GFP S2TAG Plasmid 68297 GFP Q157TAG Plasmid 68298 GFP S2TAG/E17TAG...expression vector encoding a split mCherry fluorescent reporter, enabling the detection of phosphorylation-dependent...phosphosite, mCherry fluorescence is restored, so interactions can be discovered by fluorescence-activated cell...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...
  37. CRISPR 101: Targeting RNA with Cas13a (C2c2)

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    Blog Post
    ...sequence of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in ... the RNA reporter will be cleaved resulting in activation of the fluorophore. The fluorescent signal can... a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage...programmable RNA-binding protein, creating the fluorescent fusion protein dLwaCas13a-NF for in vivo RNA ...
  38. Worm Expression Resources

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    Collection
    ... which makes them attractive for imaging and fluorescence microscopy experiments. The C. elegans genome...genome is well studied and hundreds of reporters have been developed and characterized which allow researchers...This includes CRISPR, RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids. Large ...vectors for C. elegans research, including lacZ and GFP fusion vectors. C. elegans optimized fluorophores...Approach to Better CRISPR/Cas9 Editing Efficiency Fluorescent Tagging of Endogenous Genes with SapTrap Even...
  39. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

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    Blog Post
    ...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...30220597. ACE system: A versatile in vivo fluorescent reporter for rapid quantification of base-editing...Harris lab has deposited a reporter plasmid that allows for rapid fluorescence-based quantification of base...ratio of mCherry-positive to eGFP positive cells through fluorescence microscopy or flow cytometry. Using...cobalamin-binding riboswitches that bind to cobalamin-fused fluorescent dyes; binding of the cobalamin fusion to the ...the riboswitch results in increased fluorescent signal. Prior to the development of these probes, fluorophore...fluorophore binding aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging...
  40. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...such as drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur et al., 2013), or rescue of mutant phenotypes...epitopes, such as GFP (Paix et al., 2014). A further development of this approach reported in 2017 uses the...approaches may become even more powerful following a report that one can use linear repair templates (ie. PCR-derived...templates (Farboud et al., 2019).   The Mello lab also reported that modifying the 5’ modification of oligos used...efficiencies, obtaining as many as 100 independent GFP knock-ins from a single injected animal.  PAM sequence...mixtures (Doksin et al., 2018). The Mello lab also reported efficient editing using a co-injection marker ...
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