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Showing: 331 - 360 of 1076 results

  1. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...effect of mutations of these regulatory elements on gene expression. The regulatory element (wild-type or mutant... from live cells. Monitor miRNA knockdown Expression of luciferase can be monitored as a downstream measurement...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection...Kinetics of Photon Generation Slower Faster Cofactors/Substrates Required Not Required Signal Strength...Organism Peak emission (nm) Size (kDa) Substrate Cofactors Secreted Luminous shrimp(Oplophorus gracilirostris... most often used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated...can then be used to quantify the effect of the regulatory element. Depending on the specific luciferase...

  2. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ... important, factors to consider when seeking your dream lab. The principal investigator's point of view... hires are simpler because of their documented history in science. Potential graduate students typically...goals and chat about awesome science. Important factors in the search for a lab Studies show that job satisfaction...both work style and development needs. These two factors will provide a solid foundation on which to thrive... this should (in most cases) not be the driving factor. The following questions can serve as a guide for...you take the position. Am I choosing the right mentor? Make sure your advisors will be good role models...more on this topic see our blog "Choosing a Good Mentor for Scientists". This post also includes a useful...

  3. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...bacteria that have taken up this vector, but it won’t tell you whether the vector contains the gRNA insert or...X-gal allows you to identify cells that took up the vector and distinguish between plasmids that contain the...functional 𝛽-galactosidase. Blue-white screens rely on vectors where the gene of interest is inserted into the..., you will see two bands: one representing your vector backbone and the other representing the insert....doesn’t contain the insert, you will just see the vector backbone band. Or, you can choose an enzyme that...specific to your insert or by using primers that are vector specific but amplify a potential insert. If you...plasmid does not contain the insert. If you’re using vector-specific primers, look for size differences. You...

  4. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    ...project management. Mentorship is another invaluable resource: connecting with mentors who have made similar...management often means stepping into unfamiliar territory. Unlike solving technical problems in the lab,...lab, management involves a different skill set: mentoring, setting strategic goals, and fostering team success...for me. I began exploring alternative career trajectories within the industry, but the transition wasn... role is also crucial. I owe quite a bit to my mentors, as they encouraged me to explore small-scale opportunities...company by taking on responsibilities such as mentoring a junior colleague. Guiding an intern, in particular...and leveraging tools like the Myers-Briggs Type Indicator or DISC assessments can help you navigate team...

  5. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ... your gene of interest in a plasmid and get the vector incorporated into the germline. To begin this process...knockdown. Addgene also has a number of insect vectors to consider. An easy way to see what is available...Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly There are a few important...important points to consider when choosing the vector you’d like to use to generate your new fly line:... gene to be incorporated into the genome? Some vectors incorporate into the genome randomly, while others...would you like to select for your flies? Sure, the vector will have an antibiotic resistance gene to select...transgene. One such marker is the white gene, which restores eye color to fly lines where the endogenous white...

  6. Oh, The Places You Can Go: Careers in Science Communication - Writing for a Research Institute

    Type
    Blog Post
    ...feels that storytelling is an impactful tool for science communicators. “The power of a story is to make... to learn things quickly. Story telling. The ability to know what’s a story and how to hook your readers...there’s always plenty of writing: plain-language stories about Fred Hutch research, pieces for donors such...that can be a very painful process. And certain stories can make you feel like ‘I’m a terrible writer. ...through connections and that’s something that good storytellers in science can do.” Susan’s number one piece...social media as a scientist here Learn more about storytelling in science ...

  7. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...AMA I am a strong advocate for peer mentoring, but volunteer mentors are only able to go so far with their...coach do that is different from an adviser, boss or mentor? First, professional coaches have experience and...infrastructure are not always as effective in mentoring for this transition. Second, a coach will help...experiences. Good coaches will have loads of success stories and plenty of people to vouch for them. Trust...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...online content for PhDs and postdocs posted by Toronto-based Jennifer Polk of From PhD to Life. She runs...has an outstanding blog and, while her PhD is in History, I think she would be a good match as a coach for...

  8. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...scientists who were potential Addgene requestors and depositors. China is in one of the top 10 countries...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to ...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...human tRNA genes and, until now, we've overlooked regulatory fragments of these transcripts. I remember when...was kicked off by Nobel Laureate (and Addgene depositor) Craig Mello. Taking his groundbreaking work in...Medicine presented another unexpected finding in the regulatory RNA field. The Rajewsky lab is studying circular...

  9. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...utility of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses... here! Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School ...in immune cells have identified many putative regulators of immunity. However, in vitro screens cannot... of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout ...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...responses to cancer. The screen recovered known regulators, such as Ptpn2, Pdcd1, and Zc3h12a, demonstrating...

  10. Common Injection Routes in Mice

    Type
    Blog Post
    ...Administration of substances to laboratory animals: routes of administration and factors to consider. PubMed, 50...type of injection to use depends on a number of factors. You have to weigh the pros and cons of various...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...common use of intradermal injections is studying inflammatory responses, as this method is useful for analyzing...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...

  11. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...into wide use, there were a variety of ways to monitor cell, organelle, and protein localization. For ...the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers can...Tsuboi et al.), limiting its use. Nonetheless, its creators noticed a few properties that have now become ...benefit of the common FP timers is that you can monitor age by expressing a single FP-timer protein or ... These separate proteins should be useful for monitoring cellular events occurring at varied time scales...fluorescent protein timers FP timers have been used to: Monitor dynamics of vesicle fusion and release from the...vesicle cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...

  12. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...bacteria, but have no fear - plasmids from Addgene depositors are making it easier than ever to use CRISPR  ...double stranded breaks (DSB) are lethal. Addgene depositor Luciano Marraffini’s lab took advantage of this...activation of transcription including transcription factors, phage proteins, and RNAP components (Dong et al... al., 2018). They found a transcription factor called SoxS for CRISPRa. Adding complexity, they found ...been observed with Phagemids (Selle et al., 2020, Citorik et al., 2015, Bikard et al., 2014). When the CRISPR... on the presence of specific target sequences (Citorik et al., 2014). These could be genes, specific polymorphisms...refined by the addition of a uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit...

  13. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...different Cas12a inhibitors. Some of these anti-Cas12a proteins have broad inhibitory effects on Cas12a...Caixia Gao’s lab has developed a new plant base editor from an evolved tRNA adenosine deaminase that is... is fused to a nicking Cas9. This base editor can be used for A•T to G•C edits in protoplasts or plants... Biology Find plasmids containing these base editors Spatially and temporally regulated AAV-based ...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Mobile-CRISPRi plasmids Identification of Cas12a inhibitors Using a bioinformatics and a screening approach...

  14. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

    Type
    Blog Post
    ...zinc finger nucleases to transcription activator-like effector nucleases (TALENs). Our most popular kit... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these two...now have 19 different ubiquitin plasmids in the repository. 100,000 Plasmids Shipped! - By 2010, Addgene...be popular years after they were added to the repository, including beta-catenin reporter M50 Super 8x... GCaMP6 series of genetically encoded calcium indicators were the hot technology of the year in our collection...

  15. Let There Be LITE Plasmids

    Type
    Blog Post
    ...DNA-binding domains based on transcription-activator-like effectors (TALEs) as anchors for genome engineering... “Transcriptional activators are easy to expand or combine with different effectors. I’d encourage people...The results are light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn...devised the plasmids now available in Addgene’s repository for use in neurons, both in culture and in living...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...expression levels reliably increased in vitro by a factor of 10 to 20. A flexible and modular optogenetics...

  16. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused...after that, the lab published a paper on an RNA editor that allows C to U edits (RESCUE). We’ll walk through...translational machinery treats like guanine. This RNA editor would permit point mutations in RNA which could...nonfunctional. In their quest to build a robust RNA editor, the lab started with the Cas13 scaffold, testing...PspCas13b. Find the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the...is too large to fit into adeno-associated viral vectors, they tested truncated ADARDDs to see if they could...edit multiple RNAs. It’s clear that CRISPR RNA editors will open new doors for scientific research - how...

  17. CRISPR Antimicrobials

    Type
    Blog Post
    ...papers are analogous to mammalian viral vectors in that the vectors are replication-deficient. Thus, a very...like for organisms like C. difficile. In 2014, Citorik et al. and Bikard et al. developed sequence-specific...antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, they used 1-2 CRISPR...sequences found only in certain bacteria (Figure 1). Citorik et al. targeted beta-lactam and quinolone resistance... for bacterial CRISPR targeting. Phage-derived vectors carry CRISPR RNAs (crRNAs) targeting either genomic...suggest that targeting multiple resistance/virulence factors may further reduce this likelihood. The key problem...hardest-to-kill bacteria. In using phage-derived vectors to carry CRISPR, we’re turning the tables on the...

  18. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our Blog...discovered the modular format of some transcriptional activators: that the DNA binding domain (DBD) and transcriptional...of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists took advantage... original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and the lacZ reporter gene...detectable reporter signals depend on a number of factors, including protein expression and correct folding... is to use a number of different bait and prey vectors. This has been shown to be as effective as using...transcription-activating function of a eukaryotic regulatory protein. Science. 1986; 231:699-704. PubMed PMID...

  19. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and...block out of focus fluorescence from reaching a detector or restricting fluorescence to specific excitation...light passes through a pinhole before reaching the detector. The pinhole is specifically placed to only allow...allow light from the focal plane to reach the detector and all light from above or below is excluded. ...the optics of a microscope before it reaches the detector. This means that two or more objects lying within...collected and projected back to a fast, sensitive detector such as a scientific CMOS camera. If plasma membrane...higher photon flux via the more efficient camera detector and lack of a light-reducing pinhole. By collecting...

  20. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ....com. Mentors You may have an adviser, but do you have a mentor? There is a difference. A mentor is someone...that many graduate and post-doctoral programs lack any formal or mandatory professional development training... you may need to look a bit harder to find your mentor. This may be a connection you’ve made at a conference... Alternatively, it might even be a colleague. A mentor does not necessarily need to be someone a lot older...planning, has written a great blog on choosing good mentors for scientists. Continuing professional development...popular job site webpages. Importantly, find a good mentor that can share relevant experience and resources...

  21. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...Cytosine base editors with minimized off-target DNA editing The original cytosine base editor, BE3, induces...deamination. They generated these editors using the BE4max architecture. The new editors reported in this study ...the paper in Nature Biotechnology Find the base editor plasmids CRISPRi in Caulobacter crescentus ...ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and the ctrA sgRNA...lab developed new fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of ...editing by TET1 by co-delivering TET1 and protein factors for linking methylcytosine oxidation to DNA repair...

  22. Increasing Your Work's Visibility with Addgene: Citation, Search, and Collections

    Type
    Blog Post
    ...place. As a curator of the thousands of amazing plasmid technologies contributed by our depositors, Addgene...plasmid" section Anytime you use a plasmid from the repository, we request that you cite it using both its Addgene...increases the visibility of both you and the original depositor. Our citations feature makes it so that, when ...reliability of our search results depends upon you, our depositors, giving us as much information as possible during...you to browse through the many plasmids in the repository, our scientists have curated a number of special...attention to the great variety of plasmids in the repository. And last, but certainly not least, we often ...Special Collections Pages Suggest Plasmids for the Repository Find Plasmids for Your Research  ...

  23. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ... chimeric antigen receptors (CARs). CARs are a chimera of scFvs and T cell receptors and are expressed...portion of the receptors recognizes an antigen of interest while the intracellular T cell receptor portion of...domains of the selected scFvs are inserted into a vector encoding the antibody constant domain scaffold ..., and enzymes for cancer, autoimmune, and/or inflammatory therapeutic applications. Antibody fragments...cancer-related antigens, binding of the scFv part of the receptor can activate T cells to kill cancer cells. scFv...small size also allows for their delivery by viral vectors like AAV, which could be useful for delivering ...doi.org/10.1038/nbt.1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment...

  24. Evolution of Lab Techniques

    Type
    Blog Post
    ...papers or are brainstorming new technological innovations. Let’s take a look at three historical techniques...that save both time and money. Yet, knowledge of historical techniques comes in handy whether you are perusing...real-time PCR Southern blotting, named for its inventor Edward M. Southern (1), can be used to detect ...labeling it. The label can then be visualized by autoradiography and X-ray film for radioactively labeled probes...DNA technologies that are now commonplace in laboratories. As scientific technologies rapidly progress...throughout the world. Let us know your favorite "Historical Lab Technique" in the comments section below!...Coulson. "DNA sequencing with chain-terminating inhibitors." Proceedings of the National Academy of Sciences...

  25. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    ...in the specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that...post was contributed by Magdalena Julkowska, a postdoctoral researcher at KAUST, Saudi Arabia. From the...service that we, as scientists, provide for journal editors to help decide whether work is suitable for publication... submission to the subsequent journal.  As a postdoctoral researcher studying plant development and abiotic...the data can be deposited in the public on-line repository (like GitHub, Zenodo, Figshare, Triad) while ... and Records: The Royal Society Journal of the History of Science 69.3 (2015): 337-352. PubMed PMID: 26495581...protocols Find eBooks on plasmids, CRISPR, viral vectors, and more ...

  26. The MTA at Addgene

    Type
    Blog Post
    ...therefore we are not a party (signatory) to the MTAs for plasmids in our repository. Instead, we facilitate ...and Recipient (in Addgene parlance, the depositor and requestor) organizations. Over the last twenty years...with the number of requests and deposits in our repository. From 2004 to 2023, we facilitated 784,795 MTAs...MTAs per year.  The MTA Process at Addgene As a repository, Addgene doesn’t actually own the majority of...your organization agrees to let Addgene receive, store, and distribute your material under the specified...move forward with shipping the material to the requestor.  Why the MTA?  The MTA process can be a bit confusing...

  27. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...fully-equipped laboratory for citizen scientists. In addition to maintaining a laboratory to support both... of independent Do-it-Yourself (DIY) biology laboratories working towards enabling everyone to participate... set on top of filing cabinets, a rickety old incubator, a few pipettes, and some home-made equipment....Anyone can become a member of BosLab and previous laboratory experience is not required. Our members are skilled...have non-traditional scientists working in our laboratory! Don’t worry though, we make sure that everyone... became our bacterial transformation guru and instructor for our intro to biohacking courses. Our members...a co-organizer at BosLab, a community biotech laboratory in Somerville MA. Follow her on twitter at @ZebraElement...

  28. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...licensed distributor) allow you to quickly transfer a DNA sequence from the transposon vector to one of...encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful resource....such as with signaling molecules or transcription factors, or when the gene is constitutively-active or a...edit of interest. With the larger piggyBac™ donor vector, it is possible to make the ITR and selection cassette...re-cutting. Transposase removal of the cassette later restores the target sequence. This allows researchers to...Transposase Read up on Genome Engineering with FLEx Vectors  Learn how Homology Directed Repair Is Used for...Genome Editing Resources on Addgene.org Find Vectors with the piggyBac™ Transposon Catch up on Everything...

  29. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ... DNA sequence called the Fertility factor (F-factor). The F-factor is found on an episome, a piece of ... information on viral vectors, including transduction download our Viral Vectors 101 eBook. Bacterial ...fragmented DNA released into the environment. Various factors promote natural transformation in different bacteria...bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages that attach to ...scientists commonly use phagemids, a DNA cloning vector that contains both bacteriophage and plasmid properties...

  30. How Addgene Creates Content

    Type
    Blog Post
    ...entire mission is to enable that process. Our repository of plasmids, AAVs, antibodies, and data allows...technical skills, and experiences from thousands of laboratories that makes up our Help Center, blog, website... the blog originally started as a way to easily store and share in-depth answers to frequently asked questions...on a case-by-case basis, as many of our blog contributors primarily write in the areas in which they have...graphic designer upon request that the science communicator "just sketch a western blot out quickly." Final...and cited figures from papers or image sharing repositories. Then we format, do a final review to catch ...informative resources that answer questions our depositors and customers are asking. So how can you get ...
Showing: 331 - 360 of 1076 results