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We narrowed to 863 results for: TOR;

Showing: 161 - 180 of 863 results

  1. 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." (...

  2. 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...

  3. 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...

  4. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...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 ...

  5. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...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...

  7. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...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...

  8. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...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...

  9. Addgene: The First Twenty Years

    Type
    Blog Post
    Published
    Dec. 19, 2024, 2:15 p.m.
    ...process, which was a major barrier for depositors and requestors alike. We transformed this process by ...ready-to-use viral vectors from some of our most popular lentiviral and AAV vectors, as well as developing...privilege to find and share what stories we can, and to hear the stories others have to tell.  "Also thanks...amazing years in front of us. So in our last celebratory moment, we wanted to walk through some of our...service would allow people to add a gene to a repository others could request from — Addgene!  Addgene...meant we could grow accessibility alongside our repository.  “I’m sure you hear it all the time, but Addgene...took almost six years to reach our first major repository milestone — 10,000 plasmids deposited and available...

  10. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based research as...of the gene, Cas9 activators efficiently solve these problems.   Which Cas9 activator should I use? There...plastic tool and has taken the scientific world by storm. While Cas9 has been most widely used to create ...significant work on constructing Cas9 transcriptional activators. These constructs allow for the upregulation ...no longer activated. However, there are several factors that may prevent your gene from being up-regulated... we would recommend using a cDNA overexpression vector rather than going through all the troubleshooting...genome-wide studies One of the best uses for Cas9 activators is in genetic screening. gRNAs targeting every...

  11. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...validation carried out by the commercial repository. Repositories generally ensure cell line identity and...which can be stored in liquid nitrogen long term, preferably some of these vials will be stored in a separate...in a cryostore log. Keeping good passage records allows you to track a cell line’s age and monitor any ...and money but cells lose provenance (this is the history of the cell line consisting of records of the origin...invariably much older than those available from biorepositories leading to genotypic and phenotypic artefacts...easier to conduct high quality cell culture. Biorepositories around the world ensure that the cell lines...The oldest, largest, and most diverse of these repositories is the American Type Culture Collection aka ...

  12. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...makes AAV vectors especially amenable to correcting disease-associated mutations. AAV vectors maintain...gene delivery by affinity-purified receptor-targeted viral vectors." Nature communications 6 (2015): ncomms7246... Check out our Viral Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn about Addgene AAV... is caused by mutations in the common cytokine receptor gamma chain (c) and is characterized by a lack...thereby correct the mutations using retroviral vectors. However, many diseases require techniques that...target cells has been challenging. A variety of vector delivery technologies have been investigated including...usability in clinical settings. On the other hand, vectors derived from AAV have recently gained popularity...

  13. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...are similar except that the vectors carrying the transdifferentiation factors are directly injected into... Kotton Four Factors that Differentiate the Stem Cell Field Learn how lentiviral vectors can be used for... involves overexpression of four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce a differentiated...another cell type of choice. The four reprogramming factors can be delivered and expressed in multiple somatic...supplements, bioactive small molecules, and growth factors are used to control the cell fate of iPSCs and ...         Adipocytes [28] Keratinocytes [29] Photoreceptors [30] Otic hair cells [31] Neurons [32, 33... intermediate pluripotent state is not always obligatory in cell conversion processes [35]. Rather than...

  14. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    Published
    Sept. 12, 2023, 1:15 p.m.
    ...In-Fusion cloning molar ratio calculators Lab math calculators These calculators can help you with common...lab. Molarity calculator Promega's Biomath Calculators Primer resuspension calculator (sign-in required...data repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for ....)  DailyCalcs Science Calculator app for iPhone Reagent and Data Repositories These links can help you... for plasmid design and mapping. ApE (A plasmid Editor): A free, donation-based plasmid analysis tool ...enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled...Golden Gate Reactions Cloning ligation reaction calculators These links can help you calculate how much insert...

  15. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...Stable   For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated sequences...derived. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated sequences... HB101. For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated sequences...cells for cloning and storage of unstable DNA and lentiviral/retroviral vectors. Used by Addgene for pooled... your plasmid or carrying out your experiment. History of E. coli strains E. coli are gram-negative, rod... DH5alpha and DH10b (also known as TOP10). The history of B strain is a bit more convoluted due to researchers...researchers sharing and renaming samples throughout history. It was likely isolated in 1918 but was first referred...

  16. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...reaction to generate high-throughput targeting vectors. These vectors can be used to tag endogenous genes and...the final targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9 expression...site destination vectors for inserting a tag at multiple target sites. SapTrap vectors can be easily modified...cells. To use CRISPaint, users need 3 vectors:  1) a gRNA vector that targets the gene of interest, 2)...PCR or cloning, as digestion of the destination vector with SapI yields 2 sites- the first site accepts...screening of the insertion event, and a variety of connector modules (linker sequences between the tag and ...with SapI allows the tag, selectable marker, and connector to be released (Fig. 1, Step 2-3). Since donor...

  17. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...has been exploited to localize factors such as transcriptional activators and repressors to target genes...target genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused to ... dCas9 and antibody-activator fusion proteins. The Synergistic Activation Mediator complex, generally ...promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors, and the Suntag...with CRISPR–Cas nucleases, base editors, transposases and prime editors. Nat. Biotechnol. 2020 387 38, ...array of Cas9s (many available through Addgene’s repository) and make it easy to find your perfect match....precise edit without even making a break.  Base editors are dead Cas9 (dCas9) or nCas9 enzymes fused to...

  18. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...are transfected with both vectors. Flex vectors are the solution. Flex vectors expressing fluorescent cytoplasmic.... It became possible to design FLEx vectors (Flip-excision vectors) that only allow one inversion event...concentrations of vectors expressing the required recombinase. The high concentration of FLEx vectors guarantees...experiments. Mutated FRT and loxP sites in Flex Vectors – inversion only once All recombination events ...between target sites of the same sequence in FLEx vectors, the DNA sequence that should be inverted is flanked... target sites are cleaved out, leaving behind a vector with the inverted DNA flanked by non-compatible...non-compatible target sites (Schnütgen et al. 2003). Flex vectors revolutionize neuroscience Manipulation of specific...

  19. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...imaging microscopy (FLIM) method. Acceptor photobleaching method The acceptor photobleaching method is simple...don’t measure acceptor fluorescence, this method is also less sensitive to direct acceptor excitation artifacts...interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect...fluorophore (the donor) to another fluorophore (the acceptor) by means of intermolecular long-range dipole-...dipole-dipole coupling." The emission of the acceptor fluorophore can be measured using microscopy techniques...photons) and the extinction coefficient of the acceptor (linking the quantity of absorbed light, at a ...highest quantum yield donor, the highest absorbing acceptor and fluorophores with significant overlap in their...

  20. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    Published
    Aug. 5, 2014, 6:51 p.m.
    ...responsible for storage.  If datasets are too large to transfer to a journal repository, they are likely...publishers. Nonetheless, the initial release caused a storm as the organization left open the question of how...for advice about the scope of the policy. End of story? Not quite. The PLOS policy on data access is a...statistical methods, omission of some data, etc.  The firestorm that followed implementation of the policy comprised... with limited acknowledgement of the initial contributors. This is probable to some extent - but that ...tools and assumptions to reveal findings that the originator was oblivious to, no matter how much time they...they had. It could be argued that data generators may, in light of the policy, hold onto data longer before...
Showing: 161 - 180 of 863 results