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We narrowed to 583 results for: Ret;

Showing: 81 - 100 of 583 results

  1. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    .../gamma-retrovirus Lentiviruses are a class of retrovirus (so all lentiviruses are retroviruses, but not...not all retroviruses are lentiviruses.) This means that lentivirus and gamma-retrovirus share a lot of...as tools in the lab. We will discuss the gamma-retrovirus, lentivirus, adenovirus, and adeno-associated...the common components outlined above for gamma-retroviruses. While not required for lentiviral production...   Fig. 1 - Viral vector components of retrovirus  Adeno-associated virus AAV’s elements are...are quite different from gamma-retrovirus, but they perform very similar functions to infiltrate the host...in common with AAV, just like lenti and gamma-retroviruses have a lot in common with each other. Below ...

  2. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    ...monitoring live single-cell kinase activity is FRET, but FRET reporters are challenging to design and difficult...multiplex. Problems with FRET kinase reporters Difficult to Multiplex: FRET reporters are comprised of...be multiplexed together. Challenging  to Design: FRET reporter activity depends on both the distance and...difficult to predict. Slower Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation... than KTRs which is hypothesized to be caused by FRET reporters having phosphorylated conformations that... activity.  Advantages of KTRs When compared to FRET reporters for measuring kinase activity, KTRs have...KTRs also have some disadvantages when compared to FRET reporters. First, KTRs monitor global kinase activity...

  3. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...function improperly. 3. Incorporation into a FRET biosensor A FRET biosensor is a reporter that undergoes a...include the quality control of secretory proteins in the Endoplasmic Reticulum and live cell imaging approaches...molecule are part of a single fusion protein, as in a FRET biosensor, then the local concentration of FPs around...structures including organized smooth endoplasmic reticulum (OSER) whorls. 2)A dimerizing protein of interest...potential to form large stable polymers of dimers. 3) FRET biosensors that contain two dimerizing FPs can exist...For a membrane POI localized to the endoplasmic reticulum, the normally spiderweb-like pattern of tubules...membranes termed Organized Smooth Endoplasmic Reticulum or OSER if a dimerizing FP is fused to the POI...

  4. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ... Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which a donor...acceptor. TRUPATH uses an updated version of BRET called BRET2. Figure 1: TRUPATH measures heterotrimeric...resulting in decreased BRET2 signal. With increased concentration of drug, the BRET2 signal decreases. ...excite an acceptor fluorophore. The difference for BRET is that the energy transfer is between a luminescent...bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and beta subunits... The TRUPATH BRET2 biosensor system was developed by constructing 14 luciferase donor Rluc8-Gα chimeras...emission. The activation state can be measured by a BRET2 ratio, which is the ratio of GFP2 emission to RLuc8...

  5. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...Transposable elements (TEs): Retrotransposons  Class I TEs are also known as retrotransposons. They transpose by...those without (non-LTR TEs). LTR retrotransposons are like retroviruses both in their structure and mechanism...which are required for LTR retrotransposon transposition. Non-LTR retrotransposons contain two open reading... Figure 1: Overview of retrotransposon transposition. Retrotransposons are mobilized by a ‘copy-paste...of transposition: Class I TEs, also called retrotransposons, and Class II TEs, also called DNA transposons...into new target sites. This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode...integrates in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase...

  6. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...Q1: Are retroviruses and lentiviruses the same thing? Can I package lentivirus with retroviral packaging...Lentiviruses are part of the retroviral family. What scientists often refer to as “retroviruses” are technically ...technically gamma-retroviruses - another, separate member of the retroviral family. While both lentiviruses and ...your gene of interest into cells. Unlike gamma-retroviruses that can only infect dividing cells, lentiviruses...and gamma-retroviruses use the same genes for packaging (that's gag, pol, and env), the isoforms of these...LTRs) are different. As a result, lentiviral and retroviral packaging vectors are not interchangeable. While... physiological difference between lenti- and retroviruses. Lentiviruses are able to enter an intact nuclear...

  7. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...Moloney murine leukemia virus (MMLV)-derived retroviruses. Retroviral vectors, such as pMXs or pMSCV, infect...Mechanism Efficiency Safety Pros Cons MMLV-derived Retrovirus Integrating High Low Efficient and Stable Genome...required for translational use. Viral Vectors Retroviral: Some of the first iPSCs were generated with ...generating iPSCs compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a ... corresponding endogenous pluripotency genes. Retroviral vectors are prone to incomplete silencing of ...with their differentiation potential. Finally, retroviruses are prone to insertional mutagenesis and random...integration creates variation between iPSC clones. Retroviruses are not considered safe for clinical applications...

  8. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...the best one for your experiment. Retro and lenti viruses Retroviruses are a family of viruses that stably...The most well-known retrovirus is HIV. Lentiviruses are a genus of the retroviral family that can infect...types of viruses used in mammalian research: 𝛄-retrovirus, lentivirus, adenovirus, and adeno-associated...the need for envelope disassembly. Both lenti and retro viruses can accommodate a cargo up to approximately... the host genome (non-integrating) as lenti and retro viruses do. They are generally used to achieve transient...muscular atrophy.   Type of Virus Pros Cons Retrovirus Stable Gene Expression Broad Tropism ...: PMC49545. 3. Coffin J, Hughes S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring...

  10. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...as in-stock preparations. AAV retrograde (AAVrg, also known as AAV2-retro) We’ve covered AAVrg in a previous...found to transduce skeletal muscle, cardiac, and retinal cells (Issa et al., 2023). AAV1 capsids are available...including liver, muscle, lung, CNS, kidney, and retinal cells (Issa et al., 2023; Wu, Asokan, et al., 2006...cardiac, pancreas, smooth muscle, CNS, kidney, and retinal cells (Issa et al., 2023; Wu, Asokan, et al., 2006...transduce liver, skeletal muscle, pancreatic, and retinal cells (Issa et al., 2023). AAV9 packaging plasmids... and engineered to infect neurons and undergo retrograde transport, traveling up the axon to the cell ... 2022 Pancreas AAV6, AAV8 Wang et al., 2006 Retina 7M8 Dalkara et al., 2013 Skeletal muscle MyoAAV...

  11. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ... As an example, most secreted proteins are produced in the endoplasmid reticulum, modified in the golgi...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular destination...proteins within complex structures. In order to interpret colocalization experiments in a scientific and...colocalization detected in microscopy cannot be interpreted as direct molecular interaction without further...more closely is Förster Resonance Energy Transfer, FRET. Additional opportunities: Tracking intracellular...Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka...Central PMCID: PMC3984138. Microscopy U. Basics of FRET Microscopy. Additional Resources on the Addgene...

  12. Common Injection Routes in Mice

    Type
    Blog Post
    ...tail vein or retro-orbitally Gauge: 27–30 Volume: < 0.2 ml Skill required: beginner (retro-orbital) to ...restrainer or anesthesia Tail vein or retro-orbital 27–30 < 0.2 mL Beginner (retro-orbital) to intermediate (tail...2020). A retrospective analysis for different routes of administration in Mice-Percutaneous Retro-Orbital...vein injections.” However, they can also be given retro-orbitally, behind the eye. IV injections are used...needle in the correct vein without collapsing it. Retro-orbital injections are also thought to be less stressful...between the layers of the skin. While this could theoretically be performed anywhere on the body, somewhere...researcher preferences. It sounds complicated but has a pretty low learning curve! Restraint: scruffing Location...

  13. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...et al, 2020). Because retrovirus is closely related to lentivirus, many retroviral envelope proteins have...done on enveloped viruses such as lentivirus, retrovirus, and rabies virus. How to pseudotype To pseudotype...particles.   Why pseudotype? Lentiviruses and retroviruses typically target immune cells with their wild-type...glycoprotein targets neuronal cells and can travel in a retrograde direction from the synapse to the cell body, ...allows the lentiviral vector to be transported retrograde and also has higher neuronal transduction efficiency...lentivirus with rabies glycoprotein now serves as a retrograde tracer so that scientists can identify the neurons...the replicating rabies virus envelope, travel retrogradely, and jump the synapse to the previous neuron...

  14. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...criterion than theoretical brightness for selecting the brightest fluorescent protein. Theoretical brightness...higher the number, the higher the theoretical brightness. The theoretical brightness often appears in tables...EC) and quantum yield (QY) respectively. The theoretical brightness is calculated by multiplying EC by...Cranfill et al (2016) or Thorn (2017). However, the theoretical brightness ignores several important experimental...be expected in a 'real' application than the theoretical brightness. Consequently, it makes sense to identify...Amsterdam) where she develops genetically encoded FRET based biosensors.     Additional Resources on the... theoretical brightness...

  15. AAV Viral Preps

    Type
    Collection
    ...purposes in serotypes AAV1, 2, 5, 8, 9 and retrograde Retrograde AAV To target projection neurons Caltech...

  16. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ... Looking for more? Explore our empty and EGFP retroviral plasmid set, our bacterial plasmid set for expressing... Image courtesy of IPI.   Repurposing retrotransposons for DNA editing By Emily P. Bentley For RNA-guided...beyond Cas9 with STITCHR: a combined CRISPR / retrotransposon system with the pithy name "site-specific target-primed...insertion through targeted CRISPR homing of retroelements." The Abudayyeh-Gootenberg Lab developed this...this tool based on several new site-specific retrotransposon elements they identified and characterized ...characterized (Fell et al., 2025). In retrotransposon insertion, a combined endonuclease-reverse transcriptase uses... shortened R2Tocc transposon protein, and can "retarget" the transposon to new loci in the genome. The...

  17. Structural Genomics Consortium Plasmids

    Type
    Collection
    ...AmpR 26095 pFHMSP-LIC-N N/A Expression vector for secreted proteins with N-terminal hexahistidine tag and...AmpR 26100 pFHMSP-LIC-C N/A Expression vector for secreted proteins with C-terminal hexahistidine tag and...

  18. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ... the FLIM-FRET acceptors here. See Addgene's FRET reference page for many other common FRET vectors. ...fluorescent proteins for use as FLIM-FRET acceptors  Listen to the FLIM-FRET acceptor podcast segment Article...allowing for minimized basal FRET variability, and is a good choice for FRET assays where a dark acceptor...Mork The Murakoshi lab has developed two new FLIM-FRET acceptors, ShadowG, a novel dark green fluorescent...photophysical properties and can be used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence...

  19. Editor's Choice, July 2016

    Type
    Blog Post
    ... incredibly useful in FRET experiments. Professor Fessenden's example of a FRET experiment with all relevent...into a concrete context. I recommend this post to anyone wondering if and how they can apply FRET in their...forward.   Special Delivery Fluorophore Targeting for FRET Studies While I love all of our blog posts, my pick...Hospital. This post describes how to target and use FRET flurophores for a variety of applications. While...While editing this post, I learned a lot about how FRET can be used to study biological phenomena and about...NgAgo Special Delivery: Fluorophore Targeting for FRET Studies Gendered Innovations: Why Does Sex of the...

  20. Institute for Protein Innovation

    Type
    Collection
    ...antibodies targeting families of cell surface and secreted proteins from a collaboration between Addgene ...antibodies targeting families of cell surface and secreted proteins (glypicans and integrins) and common ...
Showing: 81 - 100 of 583 results