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We narrowed to 1,075 results for: TOR

Showing: 161 - 180 of 1075 results

  1. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...designed their AAV tagging vectors (pAAV-SEPT-Acceptor and pAAV-TK-Acceptor) so that they can be easily...gene. pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR gene,...cloning them sequentially into the chosen acceptor vector. The vectors are then packaged into AAV virions via...identical targeting vectors delivered through transfection of naked DNA. Other investigators who had been working...Check out our Viral Service Find AAV Vectors Browse all Viral Vectors ...well understood, researchers have been using AAV vectors for gene editing in mammalian cells since 1998....when delivered via AAV infection, gene targeting vectors with relatively short (1 kb) homology arms underwent...

  2. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ..., Word docs, and the like is likely stored in online file storage tools like Dropbox, Box, One Drive, ...commonly used online file storage tools, as illustrated in this video, and to data stored on institutional and...source data repositories Over the past decade or so a number of general purpose repositories have been ... in the repository after the deposit has been made. It’s possible to associate the depositor’s ORCID ID...value the repository integrations, because when RSpace documents are deposited into the repositories, links...research; and (3) connectivity to open source data repositories. To illustrate this, let’s take a look at the...researchers and PIs, science librarians and data curators, IT managers, the commercialization office, the...

  3. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...communication. Viral vectors and the nervous system With the advent of viral vector technology, new methods...been used for tracing and visualizing motor, visual, and auditory neuronal circuitry in vivo. A variant... at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have been widely used...and Mooney, R. (2017). Identification of a motor to auditory pathway important for vocal learning. Nat ...genome is a single-stranded DNA molecule and AAV vectors are replication deficient (i.e. they cannot easily... low immunogenicity and cellular toxicity. AAV vectors can be equipped with strong promoters for high ... is often made Cre-dependent. If Cre-dependent vectors are used in transgenic animals that express Cre...

  4. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ... edit (or edits) engages the prime editor protein. The prime editor consists of a Cas9 nickase fused to...range of specialized prime editors (PEs). PE6a and PE6b are small prime editors with RT domains derived ...genome editing methods using nucleases and base editors have the potential to correct only a minority of...the edited strand. The first iteration of prime editor simply fused the wild-type Moloney Murine Leukemia.... We’ll summarize the major ones in Addgene’s repository later in the post. Advantages of prime editing... constrained by PAM sequence location The prime editor extends the reach of CRISPR genome editing, as ...either DNA strand, many previously developed base editors with a <8 base-pair editing window cannot edit ...

  5. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...ingredients: the lentiviral packaging vector, a transfer vector including the gene of interest, and the...3rd-generation transfer vectors apart? A3: Addgene defines 2nd- or 3rd-generation transfer vectors based on whether...between 2nd- and 3rd-generation vectors? What does this mean for transfer vectors? A4: The lentiviral packaging...largely based upon the vector system used and the transgene insert encoded by the vector." The table below ...additional information or clarification on these vectors. Read on to find the answers to our most frequently...questions. This useful resource from Addgene Depositor (and lentiviral expert) Didier Trono's lab webpage...years based on HIV-1; however, the 1st-generation vectors are never used as they pose too much of a biosafety...

  6. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ... viral vector preparations. Addgene has used this method to generally validate our viral vector preparation...of an Addgene AAV viral vector preparation after negative staining. Empty vector particles can be identified...darker than full vector particles. This image shows that the vast majority of the vectors consist of full...affected by many experimental factors. To list some complicating factors for quantifying AAV: qPCR efficiency...which is a factor in AAV titration since the starting template is the purified viral vector solution (...feasible qPCR strategy for titrating AAV vectors." Medical science monitor basic research19 (2013): 187. PubMed...many ways to quantify the composition of a viral vector solution, and generally these methods measure different...

  7. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...large size of Cas9 activators, which largely precludes delivery by adeno-associated vectors (AAV). This issue...Suntag” Cas9 activator that recruits up to twenty copies of the p65-HSF1 transcriptional activator to each ...found here. Neuron-optimized viral vectors Alternatively, Cas9 activators can be delivered to the CNS by ...mCherry (vector 1) with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed...Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center...including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches. Here...and engineered combinations of transcriptional activators to induce gene transcription at genomic sites...

  8. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...this environment produce growth factor signals such as tumor growth factor beta (TGFβ) and IL-10 that suppress... colony stimulating factor 1 (CSF-1). Additionally, tumor cells recruit regulatory T cells (T-regs) into... of other effector immune cells. TGFβ is also known to change CD4+ Th cells into regulatory cells to suppress...to inhibit expression of co-stimulatory molecules needed for T cell effector function. Cancer cells also... mice. Chimeric antigen receptors T cells producing chimeric antigen receptors (CARs) on their surfaces...contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University Cancer is one...1. Eliminate - Latent tumors are recognized by effector cells of the immune system - cytotoxic T lymphocytes...

  9. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ...and feature information given by our depositors. Many depositors also upload their own detailed plasmid...research, and helps requestors use the plasmid in their own labs. We highly encourage depositors to provide plasmid...of the Principal Investigator (PI) in whose lab the construct was made. The depositor name links to a ...starting vector that the depositing lab modified to create their new plasmid.  The Addgene Vector Database... modifying a vector previously deposited with Addgene, you can list the original vector’s plasmid number...also like to store plasmids that need extra stability, such as some of our viral vectors, in NEB Stable... used in your research! Sequence information: Depositors may provide full or partial sequence information...

  10. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    ...Agrobacterium binary vectors Vectors designed for use in Agrobacterium are termed binary vectors because they .... coli. Addgene depositor Indu Maiti has created a new and versatile binary Ti vector for both transient...pSIM24! To build a better binary vector, Sahoo et al. started with a vector commonly used by their lab, the...applications in plants. This smaller, easily customizable vector functions in multiple species, including tobacco...coli. Like plasmids used in other systems, binary vectors are cloned and prepped in E. coli. However, they...co-cultivation with the target plant species. Thus, binary vectors contain two origins of replication, one each for... be transferred to the plant, similar to viral vectors’ terminal repeats. It’s a lot of information to...

  11. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... India, and Mexico. Table 2: Addgene distributors Distributor Partnership Start Date Country Served... with a distributor. Arrows indicate the year we entered into an agreement with a distributor for that... materials in our repository. We started to increase accessibility to our repository by making it easier...Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids, both for...celebration, we thought it only fitting to share some stories of ways we’ve turned the available into the accessible...to — so far, we’ve shipped materials from our repository to an amazing 111 countries (Figure 1)! But while...would benefit from a third-party “helper”: a distributor who, among other things, could ensure our packages...

  12. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...Repressible: An activator protein is bound, turning transcription ON. When a repressor binds the activator protein...introduced the second generation QF2 activators. These activators solve the problem of broad QF expression-induced...transcription factor cannot drive transcription from the QUAS promoter, and the QF transcription factor cannot...guide to working with Drosophila Terminators and PolyA Signals FLEx Vectors Resources at Addgene.org ...the binding of RNA polymerase and transcription factors. Since the promoter region drives transcription...protein, the activator protein cannot bind the promoter sequence anymore and transcription is turned OFF....protein, the repressor protein can bind to the operator. The bound repressor then prevents transcription...

  13. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...directly to the acceptor fluorophore--think of it as a virtual photon 3. Relaxation of the acceptor fluorophore... the acceptor In order for FRET to occur several constraints must be met. The donor and acceptor fluorophores...photons) from the excited donor fluorophore to the acceptor fluorophore. The typical steps in FRET are: 1....fluorophore overlaps with the excitation spectrum of the acceptor fluorophore. Otherwise the energy transferred ... from the donor will not be able to excite the acceptor. In addition to sufficient spectral overlap, the...efficiency of FRET can be measured for a given donor-acceptor pair and a change in FRET efficiency correlates...Quantification of Förster resonance energy transfer by monitoring sensitized emission in living plant cells." (...

  14. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...RTK signaling Receptor tyrosine kinases (RTKs) are a major class of signaling receptors, but the options... for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring RTK activation...of fluorescent potassium indicators with a range of binding affinities (Torres Cabán et al. 2022): KRaION1...one of the loop regions of any G-protein-coupled receptor (GPCR) recognizing a specific ligand (Ino et al...intracellular loop of various G-protein-coupled receptors. Image adapted from Ino et al. 2022, under CC-...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...PDGFR.codonopt    Aggarwal A, et al. Glutamate indicators with improved activation kinetics and localization...

  15. 2023 and 2024: Reflections and Looking Ahead

    Type
    Blog Post
    ...into our repository, while expanding global access to our repository through our distributor partners,...developed by our depositors, used by our requestors, and supported by our repository. You can enjoy these..., a comprehensive transcription factor library, in lentiviral vectors. At 3500 plasmids, this represents... access to the Addgene repository, partially through work with our distributors in China, Korea, Japan... to ship rabies plasmids in our repository to international requestors.    I would be remiss not to mention...represents the largest single deposit in our repository and was a three–year project that touched every team...14 deposits, that became available through our repository in 2023.  In June, we announced a new partnership...

  16. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...antigens (such as viral vectors) without triggering a destructive inflammatory immune response. At the...the time, adenoviral vectors were the most commonly used viral vectors for gene delivery, and they had ...obtain FDA approval and make history in the form of Luxturna™. A better vector for ocular gene therapy: ...technician in a lab using adeno-associated viral vectors (AAVs) to treat an inherited retinal degenerative...amaurosis. This is the first of what I, scientists, doctors and patients hope are many more clinical successes... It is an easily accessible organ that can be monitored non-invasively over time. Importantly, it is an...retina but only transiently. However new viral vectors capable of providing long-term gene expression ...

  17. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector production...acids. This conjunction of vectors and exosome was given the name vexosomes (vector-exosomes), exosome AAVs...cells and photoreceptors (Wassmer et al., 2017), and exo-AAV8 expressing human coagulation factor IX in liver... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...production, assembled AAV vector particles accumulate inside the producer cells and purification protocols...purification steps. It’s also possible for AAV vectors to be spontaneously released from the producer ...Harvard Medical School took a closer look at the AAV vectors that are released from cells during production ...

  18. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...include Flp-dependent DREADDs viral vectors. Both inhibitory and excitatory DREADDS from the Gether lab are... project vectors available as ready-to-use viral preps with more on the way. S5E2 regulatory element containing...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...deployed around the world to combat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus...encompassing all possible amino acid mutations of the receptor binding domain (RBD) of the viral spike protein...angiotensin converting enzyme 2 (ACE2) cell-surface receptor. The spike protein RBD responsible for binding...2021.   After validating this method through monitoring GFP degradation, this protein compartment was...

  19. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...Cas9. Adenoviral CRISPR vectors (left) or CRISPR vectors and donor DNA vectors (right) are transduced ...need to use a two viral vectors system: one vector for Cas9/sgRNA and one vector for your specific DNA ...gRNA and Cas9 using adenoviral vectors As compared to other viral vectors, AdVs can provide a large transgene...adenoviral vectors Since then, other groups have successfully developed and used AdV vectors to edit genes...updated Dec 10, 2020 by Benoit Giquel.   Adenoviral vectors (AdVs) have been used for many years for in vivo...of oncogenicity or genotoxicity seen with other vectors (Lukashev et al., 2016). The protein capsid encapsulating...have already a packaging capacity larger than AAV vectors. This capacity is large enough to carry the Cas9...

  20. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...Subcloning - To move a gene from one vector to another simply amplify the vector and insert with separate primer...promoter of the pHLsec vector. In this scenario, IVA can be employed by cutting the vector with restriction ...García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral researcher in neurophysiology...  Javier García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...technique fail to gain traction? It is likely that both vector amplification with low fidelity polymerases and...
Showing: 161 - 180 of 1075 results