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Showing: 1 - 20 of 468 results
  1. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Updated
    Dec. 14, 2019, 3:02 a.m.
    ...AAVs) to treat an inherited retinal degenerative disease called Retinitis Pigmentosa. I quickly became...has limited tropism in the retina, primarily infecting cells of the retinal pigmented epithelium (RPE)...field of gene therapy, more specifically AAV-based retinal gene therapy. The year was 2001 and I started a...delivery, and they had been shown to to transduce the retina but only transiently. However new viral vectors...capable of providing long-term gene expression in retinal cells - AAV - were becoming available. At the same...AdV) and lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of choice ...Luxturna™, can efficiently transduce various types of retinal neurons. Nowadays a toolbox of naturally occurring...
  2. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Updated
    Jan. 4, 2021, 5:08 p.m.
    ...orientation of the FRET pair. Since many biological processes occur within the typical FRET range, FRET efficiency...undetectable FRET, well-characterized FRET reference standards can be used to validate FRET measurements... for using FRET in Your Experiments Learn about Fluorophore Targeting for FRET Studies FRET Based Biosensors...find further tips for using FRET in your experiments here.   What is this FRET you speak of? Förster Resonance...efficiency of FRET can be measured for a given donor-acceptor pair and a change in FRET efficiency correlates...for conventional light microscopy. Don't FRET - Use these FRET resources instead! As the choice of fluorophore...protein) may be able to boost FRET efficiency. Before constructing your own FRET probe, try searching PubMed...
  3. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Updated
    Jan. 23, 2020, 6:39 p.m.
    ...offense to BRET, CRET and other RET techniques which have never outclassed the Master. Note: BRET= Bioluminescence... a guitar hero (that I’m not), FRET loves playing “live.” Indeed, FRET was one of the first techniques...each other. FRET reduced the scale of our perception about molecular interactions. What is FRET? In their...Since then FRET has also been used to measure conformational changes, cleavage activity using FRET-based biosensors...its best. What are the Parameters that Affect FRET? FRET occurs when the two fluorophores used are in ...find a balance between FRET efficiency and cross-talk. Methods to Measure FRET for Cell Biology Studies...measure FRET and there is not one that is better than another. It is often recommended by FRET experts...
  4. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    Updated
    Oct. 18, 2019, 12:02 a.m.
    ...series of fluorescence resonance energy transfer (FRET)-based calcium biosensors. Perhaps most notably,... sensitivity. As reported in Nature Methods, the FRET changes of these Twitch sensor variants were optimized... can now be applied to dozens of other published FRET-based sensors. For example, his team is now applying...Technische Universiteit Eindhoven has similar interest in FRET-based sensor proteins, particularly for the intracellular... them. Merkx says, while in principle adapting a FRET sensor for monitoring calcium versus zinc should...not the other). “It’s quite a robust way to make a FRET sensor with a large dynamic range,” he says. Merkx...in the construction of redCALWY-1, a red-shifted FRET sensor for zinc using variants of mOrange2 and mCherry...
  5. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    Updated
    Nov. 22, 2019, 4:38 p.m.
    ...fluorescent units of the FRET pair together and causes an increase in the FRET signal. Subsequent demethylation...progression of cancer through different mechanisms. New FRET based tools to study epigenetics Epigenetic research...lysine-methylated peptides. This construct is in the middle of a FRET pair, which is used to visualise the results. When...separates the fluorescent units and so lowers the FRET signal back again. The Ting lab constructed 2 versions...reporter consistently showed significantly different FRET levels between unmethylated and methylated states...Fluorescent Protein Biosensors Read Our Tips for Using FRET in Your Experiments Post Read Other Blog Posts about...
  6. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Updated
    Nov. 10, 2019, 2:48 p.m.
    ...laboratory, then FRET experiments are a viable biophysical path to answers. What is FRET and what's it ...undergo FRET when they are near each other. How do you label your biomolecule of interest with FRET fluorophores...setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET and how to measure...the donor), then FRET measurements are possible. In fact, even if the majority of FRET acceptors are delivered...-targeted donors never “see” FRET acceptors, and thus never undergo FRET. If this nonspecific labeling... it's okay to be wrong In contrast to FRET donors, absolute FRET acceptor labeling specificity is not ...your FRET assay (Figure 2). These labeling methods are freely interchangeable and new uses for FRET are...
  7. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Updated
    Nov. 19, 2020, 2:15 p.m.
    ...MECP2 in Rett Syndrome Rett Syndrome was first described in 1966 by Austrian physician, Andreas Rett, who ...The Rett Syndrome Research Trust (RSRT) has an ambitious goal of curing Rett Syndrome and Addgene is ...with them to launch a Rett Syndrome Plasmid Collection and Resource Center for Rett Syndrome research. With...veteran and new Rett researchers can find the reagents and resources they need to cure Rett Syndrome.  If...If you’re new to Rett Syndrome, this blog post describes the history of Rett Syndrome research, the role...What is Rett Syndrome? Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome...neurological defects of Rett syndrome (Tillotson et al., 2017). Advancing gene therapies for Rett Syndrome MECP2...
  8. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Updated
    Dec. 14, 2019, 2:59 a.m.
    ... Viruses of the Retroviridae or Retrovirus family, which includes the gamma-retrovirus and lentivirus ...using retroviral vectors in your experiments. Gamma-retroviral vectors Popular gamma-Retroviral Transfer...genome-wide experiments. Gamma-retrovrial vs. lentiviral vector systems Gamma-retroviral vectors have a relatively...cells. Find tips on using gamma-retroviral vectors in our Gamma-retrovirus guide and find popular packaging...plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral Barcoding Library (Kit # 1000000097...viral vectors to choose from, for example gamma-retrovirus, lentivirus, Adenovirus, and Adeno-associated...enable long-term stable gene expression. Gamma-retroviral vectors are derived from the Moloney Murine Leukemia...
  9. Retrograde AAV: Making the Journey from Axon to Nucleus

    Type
    Blog Post
    Updated
    Jan. 7, 2020, 7:02 p.m.
    ...rabies) naturally exhibit retrograde transport and thus, can perform retrograde tracing, many of these viruses...exhibit relatively low retrograde transport efficiency. Because efficient retrograde access to neurons is.... In this post I’ll describe a new virus with retrograde function and how it’s enabling scientists to ...a powerful way. Keep reading to find out what retrograde function is and how it gives us better access...visualize neurons that send information to a region (retrograde tracer, Fig. 1A), and another to visualize neurons...upstream from the axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels...high-level transgene expression, but which has retrograde functionality that can be used to monitor and...
  10. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Updated
    March 12, 2020, 1:19 p.m.
    ...specificity in retinal cells. To do this, the group developed a new strategy for finding retinal cell-type ... and designed using stretches of DNA that were transcriptionally active in retinal cell types in mice....monkey, and human retinas Next, the 113 AAV promoters that were active in mouse retinas were injected into...gene expression in retinal cell types and classes in mice, primate, and human retinas. These synthetic ...types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than using naturally occurring...they drove cell type specific expression in the retina.  Four approaches for designing synthetic promoters... with sequences upstream of the start codon of retinal cell-type specific genes from mice. Strategy #2...
  11. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Updated
    Aug. 27, 2020, 1:15 p.m.
    ...Transfer (FRET). Through the non-radiative transfer for energy between neighboring fluorophores, FRET can be...function. However, FRET experiments can be difficult to implement in practice because FRET depends on a number...number of hard-to-achieve factors. For example, FRET requires that fusion proteins be in close proximity...useable data. What’s more, measuring and quantifying FRET is often easier said than done. Fortunately, a complementary...to potential protein interaction partners. Unlike FRET, BiFC only uses a single fluorescent protein to ...et al., 2002). This would be more difficult with FRET, as the overexpression of fusion proteins it requires...Addgene! How to Design a BiFC Experiment Much like FRET, BiFC experiments must be carefully optimized to...
  12. Rett Syndrome

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    Collection
    ...characteristics of Rett syndrome RettBase - MECP2 mutation database OMIM Rett Syndrome - Rett syndrome curated...Opportunities Rett Syndrome Research Trust Rettsyndrome.org References Amir et al. 1999. Rett syndrome is...Addgene's Rett Syndrome resources: Find plasmids and other research tools for use in Rett Syndrome research... Plasmid Collections Rett Syndrome Rett Syndrome Plasmid Collection and Resource Center MECP2...assemble a Rett Syndrome plasmid resource for the scientific research community. Background Rett Syndrome... mutations for Rett syndrome typically arise spontaneously, although in rare cases Rett syndrome can be... rescues neurological defects of Rett syndrome. PMID: 29019980 Return to top Animal Models Studies of ...
  13. Retrovirus Plasmids

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    Collection
    ...discuss retrovirus, they typically are referring to a subset of retroviruses called γ-retroviruses. γ-Retroviruses...FAQ) about retrovirus Popular γ-Retrovirus Envelope and Packaging Plasmids Common γ-retroviral envelope ...Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral plasmid ...Plasmids Viral Vectors 101 eBook γ-Retrovirus (gamma-retrovirus) is an RNA virus comprised of its genome and...mutagenesis). Read our γ-Retrovirus Guide for more information about: γ-Retroviral packaging systems Frequently...Retroviral packaging, envelope, and transfect plasmids Viral... Viral Plasmids γ-Retroviral Plasmids γ-Retroviral...
  14. Fluorescent Protein Guide: FRET

    Type
    Collection
    ...suitable for FRET experiments to measure protein interactions or conformation changes, as well as FRET standards...Collections Fluorescent Proteins FRET Fluorescent Protein Guide: FRET More Fluorescent Protein Resources...using the cpFRET kit from the Pertz laboratory. Empty Vectors Encoding Fluorescent Proteins for FRET The following... Back to Top FRET Reference Standards The following plasmids have well-characterized FRET efficiency values... used as FRET standards to calibrate the numerous and disparate methods used to measure FRET. Plasmid ...Background Förster resonance energy transfer, or FRET , is a process by which energy is non-radiatively...developed for use as donor and acceptor pairs in FRET experiments. The efficiency of the energy transfer...
  15. Retrograde AAV viral preps

    Type
    Collection
    ...Ready-to-use retrograde AAV available from Addgene's viral service. Retrograde construct from Alla Karpova... Viral Service AAV Retrograde AAV Viral Service: Retrograde AAV As part of our Viral Service... the rAAV2-retro helper plasmid (Addgene #81070) . This serotype permits robust retrograde access and ...viral catalog items. Browse Available Retrograde AAV See our retrograde AAV based on the functional catagories...available plasmids. Browse all Retrograde AAV Depositor Comments The AAV retro helper plasmid was generated...producing ready-to-use viral vector preparations in a retrograde serotype from the Alla Karpova and David Schaffer...viruses would be helpful to the scientific community. Return to top...
  16. TALENs for Endogenous Human Genes

    Type
    Collection
    ...TGCGGGACGTGCGGGAGCggctgcaggcgtgggaGCGCGCGTTCCGACGGCA RET TAL2346 & TAL2347 TGAGTTCTGCCGGCCGCCggctcccgcaggggccaGGGCGAAGTTGGCGCCGA...
  17. Retrovirus Guide

    Type
    Guide
    ...Glossary γ-Retroviral Packaging Systems The widespread use of γ-retroviruses (gamma-retroviruses) in cancer...pseudotyping. Retroviral Packaging Methods Return to Top Frequently Asked Questions (FAQ) about γ-Retroviral Plasmids... Overview of Retroviral Plasmid System Note : While both lentiviruses and γ-retroviruses use the same ...MoMLV and MSCV for γ-retrovirus; HIV for lentivirus). Additionally, γ-retroviruses can only transduce dividing... Are γ-retroviruses self-inactivating (SIN)? They can be. Plasmids that encode SIN γ-retrovirus lack most...Guide to retroviral packaging systems and commonly asked questions Guides... Guides γ-Retrovirus Guide γ-Retrovirus...
  18. AAV Production in HEK293 Cells

    Type
    Protocol
    ...7.0. *Pro-Tip* The pH of this solution will drift pretty rapidly upon addition of acid or base. Add only...complete media and mix. Seed all cells in 1 CF2. Return to incubator for 48 h. Trypsinize and resuspend...Seed 250 millions cells from the CF2 into 1 CF5. Return to incubator for 24 h - 36 h. Proceed with transfection...new one before putting the CF5 in the incubator. Return the CF5 to the incubator for 96 h. This incubation...minutes on ice between each pulse, 50% amplitude. Return to ice between each sonication to avoid overheating...
  19. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ... not need to obtain more competent cells and retransform. Bacteria on an LB agar plate can be stored at...prepare your plasmid DNA. In the morning, when you retrieve your liquid bacterial culture, take 500 μL of ...
  20. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...results in cell populations that are more likely to retain stable transgene expression. Other methods of generating...well in sparse/individual cultures due to lack of secreted growth factors, so not every cell type will be...
Showing: 1 - 20 of 468 results