We narrowed to 888 results for: URE

..., 2013). Two important features of this early system are its regulatable nature and its ability to multiplex...occurred. This powerful negative selection system ensures high editing efficiency; the only non-edited cells... pTargetF and pCas plasmids can be subsequently cured from the bacteria. Although developed in E. coli...Resources on the Addgene Blog Check out our CRISPR Featured Topic Page Learn more about Cpf1 Find out about...

...efficiently and accurately in a “single pot.’   Figure 1: Single-Pot Assembly: a) Graphic depiction of... sensor modules targeting a specific chromatin feature (e.g. a methylated histone such as H3K27me3), a...neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic of the interaction between the sender...quantification of this depletion using imaging-based measurements (Ashley et al., 2021).   Schematic showing...

...engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136 Perez-Pinera P, Kocak ...acetyltransferase domain capable of rearranging chromatin structure. This method has shown higher activation levels...C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional ...

...an anti-CRISPR protein. Nature 546:436–439 . https://doi.org/10.1038/nature22377 Harrington LB, Doxzen ... species of bacteria.  A note on anti-CRISPR nomenclature Acr family proteins are named for the type of... DNA cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins (Acr's) can block CRISPR...

...gene delivery, but it has a drawback: how do you ensure your gene goes where you want it to? Knowing that...transcriptionally active in retinal cell types in mice. Figure 1: The four strategies used to design synthetic...AAVs would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters can be paired for targeted...promoter AAVs in mice, non-human primates and humans. Nature Neuroscience, 22, 1345-1356. https://doi.org/10.1038...

...inheritance: 1. Genotype your parental mice In order to ensure that your genetic modification will be passed along...genotype all of your F1 mice, as well as progeny from future generations. 4. Continue breeding your colony You...they are likely to stay on separate chromosomes in future generations as well, a concept known as linkage...expression of Cre, loxP, and your gene of interest. To ensure that the effects you see in your experiment are...

...et al. "cGAL, a temperature-robust GAL4–UAS system for Caenorhabditis elegans." Nature methods 14.2 (2017... detailed information about C. elegans 3’UTRs structures, alternative polyadenylation for all protein ... novel findings. These results are normally one figure and can be negative and/or even replicated studies...

...the impacts of neural circuits on behavior. Figure 1: An overview of the chemogenetics system. ...characterize the compound, or compounds, you choose to ensure reliable results (Goutaudier et al., 2019). Using...Yang X, Stone O, Roth BL, Jin J (2015) The First Structure–Activity Relationship Studies for Designer Receptors...designed Gi-coupled receptor in transgenic mice. Nature Biotechnology 17:165–169 . https://doi.org/10.1038...

...those which you’ve already fluorescently labeled. Figure 1: Using Split-BioID to Identify Protein Protein...that give the strongest biotinylation signal, as measured via Western blot, and that result in proper localization...these two bait proteins stop interacting. This feature is particularly useful for in-depth characterization... of spatiotemporally defined protein complexes. Nature communications. PubMed PMID: 28585547. PubMed Central...

...infections. She is particularly passionate about HIV cure research and science communication.     References...to the central and peripheral nervous systems." Nature Neuroscience 20.8 (2017): 1172-1179. PubMed PMID...affinity-purified receptor-targeted viral vectors." Nature communications 6 (2015): ncomms7246. PubMed PMID... the Addgene Blog Check out our Viral Vectors Featured Topic Page Learn about CRISPR-AAV vectors Learn...

...animal’s skull using dental cement (a similar procedure as implanting a guide cannula). A fiber optic ...optogenetics but may become the standard in the future. Now that your animals are all wired up and expressing...long-term optogenetic manipulation of neural circuits. Nature protocols. 2012;7(1):12-23. doi: 10.1038/nprot.2011.413...Neuroscience research. 2011;70(1):124-7. doi: 10.1016/j.neures.2011.01.007. PubMed PMID: 21238511. 6. Jeong, ...

... explain the thinking behind the tool, its key features, how it works, and how to use it. Background on...frequency determination (CFD score) is used to measure specificity of a guide to the target. The CFD score...Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide sequences...more detailed information on using CrispyCrunch, be sure to check out the How To Guide. Conclusion CrispyCrunch...

...graduate school who were at AstraZeneca and could assure me that they were having a positive experience,... years. This time limit can feel like a lot of pressure. However, it means that your manager (and you)... choose. If publishing is important to you, make sure that your project will not involve proprietary compounds...reviewed by an internal committee before submission to ensure that it does not divulge any propriety information...

...been used to track protein localization, cell structure, intracellular trafficking, and protein turnover...plasmids to fluorescently tag markers of cellular structures in mammalian cells. These plasmids use fluorescent...reversibly degraded after the addition of auxin to the culture medium. Any number of regulatory sequences, including...“A Comprehensive Toolbox for Genome Editing in Cultured Drosophila melanogaster Cells.” G3 (Bethesda) ...

... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...drift away with each passing generation.  Another feature of balancer chromosomes is that they produce dominant... during meiosis. This can sometimes be a handy feature when setting up genetic crosses. More importantly...

...sent to the AbuGoot lab for testing, they weren’t sure what they would find… but it was safe to say the...dual activities: processing pre-CRISPR RNA into mature CRISPR RNA and cleaving RNA at positions defined.... But they weren’t done engineering quite yet – surely, they thought, the integration rate could be improved...with the single-protein CRISPR effector Cas7-11. Nature. 2021 Sep;597(7878):720-725. doi: 10.1038/s41586...

... new experimental species. Discovering ways to culture, propagate, transform, store, and share all of ... "for development of tools, approaches, and infrastructure necessary for direct tests of cause and effect...was excited to be invited to attend. My job is to ensure that, in general, these labs all include materials...from Old Faithful in 60 °C soils. Dendrobatidae (Lauren O’Connell)  Poison frogs are great for studying...

...the same time, this project gives Snapgene great exposure to the community of biological scientists who ...institutions personally on conference calls and make sure to maintain these relationships by keeping in touch...an internal to an external connection is to make sure you keep open lines of communication with colleagues...interesting events, articles and newsletters I make sure to share them with a short note - this creates a...

...stages (i.e. postdoc level) and can include the pressure to publish in high-impact journals, and the requirement...with a custom-made reagent to manipulate cells (manufactured by one of the larger biotech companies), and...company, less exposed to the fast dynamics and pressure which are often found in sales or lab-based roles...name a few). Depending on the particular company structure you may be able to move into one of these departments...

... than studies in other model organisms or cell culture. For example, Scott Lowe’s lab utilized mice to...double-stranded RNA in Caenorhabditis elegans." nature391.6669 (1998): 806. PubMed PMID: 9486653. Hesselson... and its relationship to the human genome." Nature496.7446 (2013): 498. PubMed PMID: 23594743. PubMed ...organism for studying somatic mutations and cancer." Nature Education 1.1 (2008): 183. Zuber, Johannes, et ...

...Makes Better Science Nature: What Trump’s new travel ban means for science Nature: Meet the scientists...job of having evidence”, scientists want to make sure having evidence is the job of everyone in government...all you need to do is select your state. State Legislatures: To find the website to contact representatives...

...painstakingly cloned into the perfect empty backbone, made sure to add the right tags to your gene, and may have...provide enough background for you to distinguish the features of any common lab strain and determine whether... DNA, and more. Many mutations that make these features possible are present in most commercial strains...of protein expression in E. coli and the common features found in those bacteria. Check our our blog post...

...experimental results and 2) perform targeted off-target measurements at those sites to get an idea of how high the...effects, you still need to do your due diligence to ensure they are indeed a non-issue. Above we mentioned...these outcomes. Rigorous analysis is essential to ensure off-target events are below a given threshold for...your risk level is, how to lower it, and how to ensure off-targets don’t mess with your results when they...

... either category, but those are a subject for a future post! How are repressible promoters regulated...commonly used in yeast. In the first 24 hours of culture, when glucose is abundant and ethanol concentrations...during the late ethanol consumption phase of yeast culture. Repressible Binary Systems GAL4/UAS In Drosophila...the system, researchers have used split GAL4 architecture or combined the system with Flp/FRT, as reviewed...

...contest is over, we will host other contests in the future. Subscribe to the blog to stay in the know!   ... hunt works works: You will be trying to find pictures of Addgene pets with Addgene-branded clothing ...pet gives you a clue to help you find the next picture. If you find the next correct photo, you will see...

...Addgene gave to me: a method to capture promoter-enhancer 3D chromatin structures using CRISPR. On the eleventh... you’ve been enjoying our CRISPR blog series. Be sure to subscribe to the CRISPR blog posts so you don...

...flick the tube several times. This will help ensure ensure equal distribution. Lab tip #3: Barcode everything... the latest episode.  Have some suggestions for future tips?  Send your ideas to [email protected].  Additional...

... or hydroelectric in nature. The operation of -80C and -20C freezer temperatures, biosafety cabinet ventilation... lighting/HVAC requires a significant energy expenditure here at Addgene. These are critical systems that...

...What's a lab group in industry? The collaborative nature of working in industry and how delegating can empower...success. The importance of thinking about the big picture and not just the next experiment. Chapter 6: Advice... the treatment of various diseases, from heart failure to Alzheimer’s.  You can follow her on Twitter ...

...29:54 - 31:25 : Nature's defenses against gene drives. 31:26 - 33:08 : Plans for future work at the Wyss...advisory board where he will help shape Addgene's future. Listen to this episode for insights into the process...

...key step to any experiment and can help prevent future headaches. To help you plan PEI transfections for...simple to make adjustments for the type of tissue culture dish and number of dishes used. Here’s to making... pHelper: 2 RepCap Select what type of tissue culture dish you will be performing the transfection in...

...for years – searching our photo archives I found pictures back through 2009, though I suspect that our costume...months in advance. Our teams comb the news (pop culture events of the past year obviously make for good...takes home the ultimate prize – a glorious trophy. Sure it’s plastic and you have to share 1 trophy among...

...what the founders hope to see in the repository’s future. If you’ve ever been curious about what it takes...success Chapter 6 - Developing Addgene’s Brand and culture Chapter 7 - Building Addgene as a global organization...Tufts Launchpad, and more Chapter 10 - Addgene's Future, the research team, and the importance of sharing...

...perform in vivo screens in other cell types. A key feature of this system is a lentiviral gRNA expression ...the statistical power of in vivo screens. Figure 1: Using FITS to perform in vivo CRISPR screens...clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission from Milling and Markson et...

...and look forward to hearing from her in the future. Future awardees We will be opening the Addgene/Seeding...them the most. Check back here to learn more about future awardees and their inspiring research. Additional...

...and proteins associated with pathogenesis. In his future work. Dr. Bengyella would like to use the expertise...to do excellent work in the biological sciences. Future awardees Now that this first round of awards has...enable scientists in developing regions in the near future. Additional Resources Apply to Recieve Equipment...

...also be expanded to include even more loci. Figure 1: Schematic overview of the all-in-one CRISPR/... work successfully to target multiple sites in cultured human cells. While it has not yet been demonstrated...dCas9-KRAB in our system,” Sakuma said. “These are sure to be powerful tools to modify the expression of...

...Today’s episode of the Addgene Podcast features guest host and Addgene European Outreach Scientist, Benoit...cooperation, and antibiotic production. Chapter 5: The future of Synthetic Biology. Wide adoption of synthetic...Biology and Innovation, Imperial College London and lectures synthetic biology for the Department of Bioengineering...

...University). Right there on the floor outside the lecture hall. It was wonderful to see the ad-hoc ‘sciencing...in vivo using ultrasound (for an example see the figure below). In addition to low costs, deep tissue penetration... imaging of microorganisms in mammalian hosts." Nature 553.7686 (2018): 86.PubMed PMID: 29300010. PubMed...

...more secrets - during a collaboration, members of Maureen Hanson’s and Rima Menassa's labs made the accidental...nm, photoconverted with 405 nm light, and then captured in the red state after excitation at 561 nm. This...conversion did not occur. Tobacco suspension cell culture with cytosol-targeted GFP. White circles represent...

...substantial amount of properly folded, relatively pure protein, but getting to this stage is often much... coli strains designed for protein expression. Cultures are then lysed and the protein of interest is ...disulfide bond formation. pCri also includes a special feature for difficult-to-purify membrane proteins. pCri...

...products is desired, as multiple selection markers can ensure that all desired tags have been integrated. Simply... to their genes of interest, allowing for easy capture and detection of proteins and complexes, without... single strain. In addition to the collections featured above, many other modular yeast tagging systems...

...progress towards understanding this disease. Figure 1: Scientists are continuing to do essential research...portal has recruited 3,218 Canadian volunteers, and features over 200 COVID-19 related research projects, funding... will remain an extremely important part of the future of research. To find other COVID-19 databases and...

...active in both E. coli and human cells. Using the structure of CasX at eight different states of assembly .... coli and human expression. Read the paper in Nature Find the CasX plasmids Drug inducible transcriptional...sequence-specific toxicity of Cas9. Read the paper in Nature Communications Find the Bikard E. coli Genome-wide...

...scientist in training can adopt to ensure opportunities in the future. Studies show that the majority of...an excellent resume ready to be submitted The structure and content of a resume can be crucial for a successful...

...proteins known to localize to specific cellular structures.The Davidson lab even included some of their own... see Supplemental Documents here). If you’re not sure where your protein of interest localizes in the ...easier for scientists everywhere to get a clearer picture of that intracellular world, even as the Davidson...

...This blog post is the first in a series that will feature advice for students, postdocs, and young Principal...their top 3 tips. Tom Ellis 1. Keep up with the literature. He recommends setting up a few Pubmed email ...“If you don't read, you're doomed.” 2. Always be sure to include various positive and negative controls...

...paper that appeared in Nature Biotechnology back in 2011 and then in another Nature Methods paper last year... far-red light. As the researchers described in Nature Communications last year, “The capability to control...

...archaea and eukaryotes such as yeast and plants. In nature, plasmids provide one or more functional benefits...for the life scientist or bioengineer.   Figure 1: Map of a plasmid with its elements described...constructed in the lab? Due to their artificial nature, lab plasmids are commonly referred to as “vectors...

...get it into your plasmid backbone, and zap the mixture of DNA into cells. Unfortunately, not every cell...post, but don’t worry, we’ll cover screens in a future post. Positive selection: Cells that have gained...toxin genes, and the sacB gene). By tapping into nature’s toolbox, scientists are finding more and more ...