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We narrowed to 116 results for: GFP reporter gene

Showing: 41 - 80 of 116 results
  1. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    June 6, 2019, 12:41 p.m.
    ... developed four acoustic reporter gene (ARG) plasmids to better monitor gene expression in vivo using ...plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive...contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years, scientists ...rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue performance...demonstrates engineered gas vesicles as the first reporter genes using ultrasound and provide a major benefit...29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated gene regulation Article...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into...
  2. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

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    March 5, 2019, 1:08 p.m.
    ...deletion of one or several of these genes permits the expression of genes farther downstream in the construct... two tandem sets of genes aligned head-to-head and the deletion of one of the gene pairs. Brainbow 2.0... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ...recombination in this mouse generates cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic...Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent proteins are organized ...organized in tandem with a single promoter, but only the gene directly following the promoter is expressed. Cre-mediated... which uses Cre to trigger the inversion of two genes aligned head-to-head. Another variation, Brainbow...
  3. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

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    Blog Post
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    Aug. 15, 2017, 1:24 p.m.
    ... to track where GFP labelled B cells were in mounted slices of these organs. These GFP labelled cells ...enabled by fluorescent proteins The optimization of GFP and its derivatives by the late Roger Tsien changed...stimulus. Using an epifluorescence microscope and GFP fusion proteins controlled by promoters specific ...the immune lineage, scientists could easily track GFP expressing cells. For example, by harvesting lymph...transgenic mice expressing fluorescent protein reporter constructs in the immune lineage. By specifically... the infiltration of neutrophils expressing LysM-GFP (green) into a draining lymph node after infection...using 2-photon microscopy as this hasn't been well reported for many fluorescent proteins. Additional fluorescent...
  4. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    Blog Post
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    April 9, 2019, 1:13 p.m.
    ...the limitations is the size of GFP: ~240 amino acids or about 28 kDa. GFP and its homologues have a beta-barrel...fluorescent protein (GFP) from Aequorea victoria and its variants are genetically encoded fluorescent ...interference is known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites...post-translational modifications. Despite these potential issues, GFP has been used successfully in countless fusion proteins... an inert light emitting module, any tendency of GFP to homodimerize should be avoided because it will...the linker sequence short In the early days of GFP applications, many were concerned with steric hindrance...fluorescent proteins derived from Aequorea victoria GFP sticks out and can be considered a linker (figure...
  5. Fluorescent Proteins 101: Fluorescent Biosensors

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    May 18, 2017, 2:30 p.m.
    ...molecule it detects. This change then generates a reporter signal. Reporter signals may be electrochemical or...biosensors To focus on a particular gene or protein, check out our gene-specific biosensors. These constructs...light-based, with luminescent and fluorescent reporters being especially popular. We’ll give you an introduction...biosensors as an example. Many calcium biosensors include GFP fused to calmodulin (CaM) and the chicken myosin ...glutamate sensor pCMV(MinDis).iGluSnFR. Picking your reporter Don’t forget about the other end of the biosensor...Visualization Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent... OR enzyme activity. Small molecule biosensors Addgene has many sensors that measure the concentration...
  6. Hot Plasmids and Viral Preps - September 2021

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    Sept. 21, 2021, 1:15 p.m.
    ...systems allow researchers to understand bacterial gene clusters that produce useful natural materials, ...capable of robust expression of high G+C content genes, as validated through monitoring the expression ...plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE, to identify...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...through the success of the change in expression of GFP, for the stop codon should have changed from “TGA...cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s. b) Quantification ...human mRNA binding proteins with 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode...
  7. Popular Retroviral Vectors and Their Uses in Scientific Research

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    Blog Post
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    July 24, 2018, 12:51 p.m.
    ...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...
  8. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
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    Dec. 17, 2020, 1:30 p.m.
    ...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...
  9. Hot Plasmids - August 2020

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    Aug. 18, 2020, 1:15 p.m.
    ...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...
  10. 10 Great Guest Posts We're Thankful For

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    Blog Post
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    Nov. 18, 2016, 3 p.m.
    ...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 (...
  11. Fluorescent Proteins 101: Aptamer Fluorophores

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    Blog Post
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    April 11, 2017, 2:30 p.m.
    ...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...
  12. New Viral Vectors - Fall 2024

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    Blog Post
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    Oct. 3, 2024, 1:15 p.m.
    ...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...
  13. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    April 25, 2017, 2:30 p.m.
    ...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...
  14. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
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    Oct. 22, 2015, 2:30 p.m.
    ...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...
  15. Another Pathway into Cells: iTOP

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    June 23, 2015, 8:37 p.m.
    ...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...
  16. Negative Can Be Positive: Open AAV Data with Addgene

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    Blog Post
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    Sept. 6, 2022, 1:15 p.m.
    ...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–...
  17. Choosing Your Fluorescent Proteins for Multi-Color Imaging

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    Blog Post
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    Oct. 9, 2014, 3 p.m.
    ...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...
  18. Plasmids 101: Mammalian Vectors

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    Blog Post
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    March 25, 2014, 3:15 p.m.
    ...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...
  19. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

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    Blog Post
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    June 4, 2019, 1:02 p.m.
    ...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...
  20. Hot Plasmids: Fall 2025

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    Blog Post
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    Nov. 11, 2025, 2:14 p.m.
    ...fused to GFP forms fluorescent biomolecular condensates. A GFP-nanobody fused to the killswitch (GFP-nb–KS... killswitch can be genetically fused to a protein of interest or targeted to a GFP fusion protein using...Grelet Lab developed MitoTRACER, a retroviral-based reporter system that permanently labels the cells receiving...using an anti-GFP nanobody (Figure 4). Once incorporated into a condensate, the killswitch disrupts its ...CRISPRa components to adipose cells to activate genes involved in converting white adipose tissue (WAT... metabolizing glucose to dissipate heat. Of the genes they tested, activating UCP1 had the most potent...increasing oxygen consumption and upregulating other genes normally expressed in BAT. When donor-derived CRISPRa-AAV-treated...
  21. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
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    June 30, 2015, 2 p.m.
    ...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 ...
  22. Hot Plasmids: Summer 2024

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    Blog Post
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    Aug. 27, 2024, 1:15 p.m.
    ...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...
  23. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    Blog Post
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    June 17, 2021, 1:17 p.m.
    ...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...
  24. Choosing Your Perfect Empty Backbone

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    Blog Post
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    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...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...
  25. Viral Vectors 101: Calcium Sensors

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    Blog Post
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    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...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...
  26. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Sept. 4, 2018, 12:53 p.m.
    ...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...
  27. Hot Plasmids - February 2022

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    Feb. 22, 2022, 2:15 p.m.
    ...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...
  28. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

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    March 3, 2020, 2:15 p.m.
    ...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...
  29. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
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    Feb. 8, 2018, 12:17 p.m.
    ...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...
  30. Lighting Up Cell Signaling with Photoswitchable Kinases

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    Blog Post
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    July 25, 2017, 1:20 p.m.
    ...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...
  31. pSiM24: Simplifying Plant Genetic Engineering

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    Blog Post
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    Sept. 29, 2015, 2:30 p.m.
    ...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...
  32. Simplify Cloning with in vivo Assembly

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    Blog Post
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    Oct. 18, 2018, 12:37 p.m.
    ..., 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...
  33. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    Blog Post
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    Aug. 5, 2021, 12:15 p.m.
    ... 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. Plasmids 101: What is a plasmid?

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    Blog Post
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    April 2, 2020, 6:17 p.m.
    ...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 ...
  35. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    Blog Post
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    Aug. 11, 2020, 1:15 p.m.
    ...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 ...
  36. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    Blog Post
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    Dec. 13, 2018, 1:14 p.m.
    ...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...
  37. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
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    Dec. 6, 2022, 2:15 p.m.
    ...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...
  38. New Viral Vectors - Winter 2025

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    Blog Post
    Published
    Jan. 28, 2025, noon
    ...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...
  39. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...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 ...
  40. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...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...
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