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We narrowed to 1,075 results for: TOR

Showing: 221 - 240 of 1075 results

  1. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...efficiencies of integrating viral vectors. New Sleeping Beauty transposase vectors Even more recently, Eric ...developed two new types of vectors for the SB transposase system - one set of vectors for robust constitutive...transposase as a nonviral vector for CD19-directed chimeric antigen receptor (CAR) therapy. SB is a valuable...of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both introduce...cells’ defense mechanisms. The use of non-viral vectors, like transposons, avoids many, though not all,...use of viruses is also important for social and regulatory reasons. 1.) The transposon consists of a mirrored... expression (pSBbi derivatives) and one set of vectors for inducible expression (pSBtet derivatives). ...

  2. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...combinations of promoters, terminators, solubility and purification tags, connectors, cDNAs, and more to find...site-specific recombinases, regulatory elements, developmental regulators, and lots of viral components...workhorse yeast — K. phaffii (formerly known as Pichia pastoris). If your goal is protein isolation, then the ...kit has a focus on infrastructure with shuttle vectors, several delivery systems which can be engineered...engineered into the TUs, and even viral gene silencing vectors. The Plant Parts III kit has a focus on transformation... examine the effects of different promoters, terminators, enhancers, and cross-cassette interactions in...effects of protein degradation on chimeric antigen receptor signaling (Kim et al., 2024). The EMMA Toolkit...

  3. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...chair of the postdoctoral engagement committee and active member of the university's Postdoctoral Association...science on April 22, 2017. While experiments (and editors) can move slowly, news about possible broad changes...in the veracity of its work. Alina Chan is a postdoctoral researcher at Harvard Medical School working...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...health depends on it. Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072...to join the public conversation and to tell the stories behind our research.  That's why I marched for ...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...

  4. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    ...about being an editor - that I learned on the job. The day-to-day work of a science editor In my role as...sorts of different editors. Do you like science journalism? Take a look at our news editors. A fondness for...at our multimedia producer and online editor. And, the copyeditors deal with things like word choice, punctuation...contributed by guest writer Pamela J. Hines PhD, Senior Editor at Science Magazine. Although we only walk one ...be like? Then I spotted an opening for being an editor at Science magazine. That one I just could not ...as an editor at Science, my various interests come together as useful and productive, forming a different...science journalists, or policy makers. The term editor covers a wide variety of careers and jobs. At Science...

  5. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...employing optogenetic regulators to initiate the event being monitored. Optogenetic regulators respond to specific...fluorescent protein (acceptor) in the pair. The higher the emission from the acceptor, the more powerful...and NanoLuc® Luciferase are available from the repository. We encourage people to go to Addgene to get ...furimazine by Promega’s Chemistry group is a good story (1). From a deep sea shrimp to a compact powerhouse...applications of the Enhanced Nano-Lanterns including monitoring multiple cellular events such as the dynamics...initiation wavelengths, excitation of the optogenetic regulator leads to spurious activation of the FRET sensor...still transfer energy to the fluorescent protein acceptor. NLuc is an excellent BRET donor due to its bright...

  6. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...might not be a big factor for you. Know who's boss You sign on to a lab for the mentoring only to find out...choose a mentor that sees you only as a paper making machine. Choose one that is a true mentor, supporting...rotation was with a mid-sized lab with an experienced mentor. The lab was very, very quiet. Every time I asked...some of these questions in the sections below. Mentoring relationships can help identify your priorities...not worked for you in the past. For all of the factors I touch on below, the first question to ask yourself...“which ones matter to me?”  and hone in on the factors that will make the most difference for your success...? Will a postdoc or lab manager be your actual mentor? Is there a chance the supervisor will be moving...

  7. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...him a more effective motivator (“be like Tony Robbins!” he encourages faculty mentors.)  Collaboration Maximizes...technical and pedagogical resources, offers mentoring for instructors, and provides opportunities for students...in a year-long biochemistry laboratory : Part II – research-based laboratory – a semester-long research...becoming just the fifth repeat Nobel Prize winner in history. Interestingly, he also adds to the disproportionately...this trend as evidence that the best teachers and mentors pass on more than just technical knowledge, shaping...responsibilities left him with limited time for one-on-one mentoring. His solution? Bringing his research into the ...work identifying new genes required for stem and progenitor cell maintenance in planarians in Developmental...

  8. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...has a great team of moderators who facilitated scheduling, filtered out inflammatory questions, and even...and the moderators were on top of any comments that were at all trollish. Kudos to the moderators for keeping...trigger phrase these days). Redditors ask serious questions What the moderators did not delete or suppress...incredibly important to find good mentors and incentivize good mentorship: AMA mechanics: Be prepared! The...Addgene Executive director, Joanne Kamens, recently participated in a Reddit AMA (short for “Ask Me Anything...lists of forums on particular topics. Volunteer moderators set the rules for these forums and try to keep...careers, women in STEM, work-life negotiation, and mentorship, we decided to have her participate in an AMA...

  9. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...empty control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows... In this context, the empty vector control shows any effect of the vector/backbone itself on gene expression...hypothesis: proper controls! For every experiment, an investigator needs a standard against which the results can...variable you are testing and not some other unintended factor. Control plasmids are used to minimize any effects...planning your experiment includes determining what factors need to be controlled for in order to eliminate...replicate controls.  Transfection controls: empty vector and internal control A transfection control measures...any effects of the transfection itself (i.e., the vector used in transfection, transfection reagent, or ...

  10. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...few inventory items offered domestically, but by the end of 2016, we expanded our viral inventory to 25... is a dual-vector system, meaning it is typically used in conjunction with a second vector that delivers...constructs (i.e., using a dual-vector system as opposed to a single-vector system) has several benefits.... lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide...LentiCas9-blast LentiCas9-blast is a Cas9 expression vector developed by Feng Zhang’s lab at the Broad Institute...such as lentiGuide-Puro. The alternative single-vector systems typically express Cas9 and a gRNA from ...compared to separate transductions with a single-vector system). Tips for using this plasmid Cas9 expression...

  11. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ... Set up a table of contents in the top directory: A directory is a file system in the computer and is...experiment its own directory: In addition to output files, this is also where you can store any power point...in C++ and Java Command line history: Tracking command line history by recording all commands entered...analysis like a wet lab experiment I use the text editor TextWrangler to record the following, which are...name for a folder. For instance, your Desktop directory contains the folders and files that you see on...on your desktop. The location of your master directory or folder for all of your analyses is up to you...README.txt document is invaluable. In the top directory of every experiment, immediately write a text ...

  12. The time and cost required to make a plasmid

    Type
    Blog Post
    ...restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise the...use of both when making plasmids. Use repositories Repositories such as Addgene help share materials quickly...After the great Plasmid Debate of ’18, Blue Flame depositor John Doench calculated the time to make a plasmid...for the simple insertion of a PCR product into a vector. The time it takes to make a plasmid could increase.... This cost varies hugely based on a number of factors, but we can assume it’s at least equal to the $155...in a 96-well (or 384-well) plate! How much do repositories help?   Designing precise calculations for any...any of the numbers in this post is a mathematical story well beyond my ability to tell. But even from these...

  13. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little while Transient...? Check out our Retro expression vector and Lenti expression vector. Turn it on for just a little while...CRISPR for that, too! CRISPR activators (CRISPRa) are transcriptional activators fused to a dead Cas9. The...out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the time If ...Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging vector! Destabilizing domains Destabilizing...increase expression. There are several flavors of activators which can be used with this approach - read more...them on our CRISPRa blog! Fig. 1 CRISPR activator and repressor mechanisms Want to transiently...

  14. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases, and more. With ...by using cis regulatory elements such as promoters and enhancers, multicistronic vectors, and post-translational...Addgene blog Download the viral vectors 101 eBook Read our viral vector related blog posts Learn more about...With the help of adeno-associated viral vectors (AAV), neuroscientists can combine classical genetic tools... how these changes affect the phenotype. Viral vectors and genetic approaches are continuously refined...Education and Development workshop aimed at brainstorming strategies for using combinations of AAV-based...designing experiments with AAV: 1. Getting your viral vector into your cell of interest: AAV tropism and route...

  15. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ... they monitor. Gene traps Gene traps are promotor-less constructs that contain splice acceptors. They...below), but fundamentally the reporter construct monitors the expression of a nearby gene. The reporter ...high/low) and patterns (tissue/timing) can be monitored under various conditions without necessarily modifying...stage you could infer that the gene which GFP was monitoring was being expressed in the same pattern. These...throughout an organism’s genome. These reporters have historically been used extensively in fly, mouse, and yeast...result but will still allow for expression pattern monitoring. Enhancer traps Like promoter traps, enhancer...traps, it will still likely disrupt the gene it monitors due to transcriptional fusion.   Examples...

  16. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ... of stimulation to receptors. Optogenetics offers better control over these factors as the light source...more about optogenetics New to viral vectors? Browse our Viral Vectors 101 blog posts Learn more about precise...Addgene.org Browse all viral vectors available at Addgene Visit our viral vectors guide Find our chemogenetics...different advantages and limitations.  The four main factors in which chemogenetics and optogenetics differ ...offers a high degree of temporal resolution as receptors can be turned on and off simply with the presence...diffusion of drugs throughout the body, the onset of receptor activation takes minutes. As a result, it is not...response or track the exact time of activation. Receptor activity is also drawn out as drugs can remain...

  17. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...inheritance. CRISPR activators and repressors are thought to be reversible once the effector is inactivated...promoters. Other researchers additionally fused a TAL effector to LSD1 histone demethylase to demethylate enhancer...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, is available...and repair pathways work downstream of Tet1 to restore the DNA after cytosine removal. Recently, Albert...alone, or coupled with DNA oxidation and repair factors that allow for increased gene activation at the... targeted enhancers. Although the lab used this vector in chick embryos, it also functions in mammalian...and this fusion allows for packaging into viral vectors and targeted delivery into hard-to-manipulate cell...

  18. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...bacterial prey. Currently, receptor binding proteins (RBPs, the host recognition factors of phages) constitute...Therefore, in addition to their utility in the health sector, phages represent a reservoir of carbohydrate binding...function of digestion time. For phage DNA that is refractory to enzymatic digestion, however, a physical fragmentation...a hose to a nitrogen gas tank with a pressure regulator. Other methods for physical shearing exist as ... blunt ends) and ligate into a bluntly digested vector designed for protein overexpression in E. coli,..., such as pET30a. If working with DNA that is refractory to enzymatic manipulation, amplifying phage DNA...primers bordering the multiple cloning site of the vector. In order to reduce the chances of some clones ...

  19. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...Louis provides a combinatorial indexing strategy that simultaneously tracks clonal history and cell identity...embryonic fibroblasts (MEFs) into induced endoderm progenitors (iEPs), a self-renewing cell type that can differentiate...iEPs by retroviral expression of transcription factors Foxa1 and Hnf4α, a process that like other reprogramming...CellTagMEF), at the end of the three-day transcription factor reprogramming treatment (CellTagD3), and 13 days... Morris Lab found that cells took one of two trajectories: a route towards successful iEP conversion or... The CellTags revealed that cells with common ancestors tended to have similar reprogramming outcomes....2019). Key takeaways of CellTag CellTag is a combinatory cell-indexing tool that combines scRNA-seq and...

  20. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ...Existing methods for AAV quality control Historically, AAV vector QC has been sparse including only titration... procedures and Python scripts with laboratories and viral vector cores routinely producing AAV. We believe... our viral vectors related blog posts Read our blog post on AAV titers Browse viral vectors protocols ... platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process,... of quality control is sufficient but for viral vector core facilities producing a large number of viral...demonstrated that NGS is an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing... simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly and...
Showing: 221 - 240 of 1075 results