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We narrowed to 854 results for: TOR

Showing: 141 - 160 of 854 results

  1. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type IIS restriction...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...-VP64 (transcriptional activator)-containing plasmids. Each destination vector contains GFP, enabling ...create lentiviral vectors expressing wt humanized Cas9 and up to three gRNAs. Entry vectors containing the...wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing other...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...Joachim, et al. “A versatile modular vector system for rapid combinatorial mammalian genetics.” Journal of ...

  2. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...students, and those from historically underrepresented groups. Through such mentoring programs, scientists...especially grateful to its mentors and supporters during this first year. Mentors Amanda Gunn, Bryan Gibb.... The teaching/mentoring bug never went away. While he has frequent bouts with Impostor Syndrome, Joe ... done at my first six ASM meetings. I told this story over a great dinner at Pittypat’s Porch with old...James et al., 2020. Help me stranger - Finding mentors for the meeting Just like analysis pipelines for... meetings. A key was to provide each bin with a mentor who had attended the meeting multiple times before...plan for and network at the meeting. We found our mentors by asking for help with the program on social media...

  3. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...vivo studies. Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by...to determine which facility produced your viral vector lot. At Addgene, AAV particles are titered by droplet...method.  Titering by the University of Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR...

  4. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...community concerns AAV vectors are complex biological reagents; many factors, like storage buffers and conditions...when I started working with AAV vectors. I wanted to use an AAV vector to express ChR2(H134R) in A2 norepinephrine...to AAV vectors, many from people trying to answer the exact same question I was: What AAV vector is best...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists...question and used it to publish!  Working with AAV vectors in neuroscience Let’s consider my first challenge...didn’t provide any information on other important factors such as what is the best promoter to use, or if...deal of time and effort to test and optimize our vector - overall, I spent two years testing and reformulating...

  5. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    Published
    Oct. 22, 2014, 1:57 p.m.
    ...Choose a moderator with a backbone. It is nice to have a subject area expert, but the moderator is not ...LinkedIn profiles of all panelists to the moderator. The moderator and organizers should develop a list of...interviews, the moderator should be taking good notes and discovering interesting stories, riffs or opinions...contribute. The moderator should identify the funny, provocative or interesting stories that will make ...bring out these interesting stories and remind the panelists of the stories or topics you know they can...(lapel mics are best). The moderator takes control It is the moderator's job to prevent anyone from ...published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates. Panel discussions are becoming...

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

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ...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...

  7. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    Published
    Jan. 17, 2019, 1:38 p.m.
    ..., 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...

  8. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ... 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 ...

  10. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ... 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...

  12. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...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...

  13. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...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...

  14. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ...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...

  15. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    Published
    Sept. 29, 2015, 2:30 p.m.
    ...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...

  16. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ... 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...

  17. Plasmids 101: Repressible Promoters

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

  18. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...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." (...

  19. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...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...

  20. 2023 and 2024: Reflections and Looking Ahead

    Type
    Blog Post
    Published
    Jan. 23, 2024, 2:15 p.m.
    ...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...
Showing: 141 - 160 of 854 results