...between the purine bases (adenine, guanine, and hypoxanthine) or between the pyrimidine bases (cytosine, ... purine to a pyrimidine or vice-versa. Note: hypoxanthine (shown) is the nucleobase component of the nucleoside...have scientists solved this problem? By taking advantage of repair pathways within the cell — in some cases...Created with BioRender.com. Base editors take advantage of this single-stranded DNA (ssDNA) region, using...they expanded CGBE target scope by creating BE variants with other Cas9s that recognize different PAMs...processes that would cause DSBs, and its inclusion significantly reduced indels generated by CGBEs. What's next... aspiring genetic editors can now compare the advantages of base editing and prime editing, an alternative...

...tell only part of the story. This year, we made significant strides in making science more accessible and...serotypes, or hundreds of Data Hub reports detailing antibody experimental results. We also redesigned our website...growing collection of thoroughly characterized antibodies, and our contributions to key research papers...papers are already inspiring higher standards in antibody use (Ayoubi et al., 2024; Kahn et al., 2024). Building...materials — we're accelerating the future of science. Want to learn more about our latest developments? Subscribe... Laflamme, C. (2024). A consensus platform for antibody characterization. Nature Protocols. https://doi.org..., A. R. M., Cable, H., & Longworth, S. (2024). Antibody characterization is critical to enhance reproducibility...

...proteins (FPs) are also the foundation of multiple important technologies, including FRET and optogenetics....This simple technique has been shown to work in plant, Drosophila and mammalian cells, and it may find...was made by accident. After exciting GFP-labeled plant cells at 405 nm instead of 488 nm, Sattarzadeh et...imaging process worked well with the common GFP variants EGFP, mGFP4-T, and S65T GFP, and no red color ...can be rapidly and efficiently photoconverted in plant, Drosophila, and mammalian cells. Photoconverted...enthusiasts! Addgene has empty backbones containing GFP variants used by Sattarzadeh et al., as well as a wide ...Addgene’s Fluorescent Protein Plasmids and Resources Relevant Blog Posts Plasmids 101: Green Fluorescent Protein...

...through the ‘front door,’ the homepage is still important. It needs to express value, clearly communicate... what a site offers to their users, and most importantly, allow those users the ability to quickly access... to improve search and browse functionality. We wanted users to be able to find plasmids and viral vector...Along with improving search and navigation, we also wanted to emphasize the values that make Addgene unique...and allowed our content to dictate the space. We wanted to comfortably tell our story, to get scientists...500th time on addgene.org. Additionally we also wanted to highlight our blog, daily updates, and offer...conferences, on our website, or in social media, we wanted to make sure our friend matched our core branding...

...alpha2-AR antagonist, nLight sensors are not. Instead, they are blocked by alpha1-AR antagonist. This is...Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been difficult...is important because alpha2-AR can provide inhibitory feedback in response to norepinephrine release, ...the nLight indicators. Speed One of the major advantages of this next generation of norepinephrine sensors...GRABNE, with both nLightG and nLightR showing significantly increased selectivity for norepinephrine over.... In particular, they’re focused on generating variants with improved affinity and dynamic range of both...

... and antibody labelling. In these experiments, biotin is added to a primary or secondary antibody that...organisms for life. Biotin is only synthesized by plants, most bacteria and some fungi, thus mammals must...of carboxyl groups to organic acids to produce important cellular metabolites. Biotin acts as a carboxyl...that targets your protein of interest or primary antibody respectively. Avidin bound to a reporter is then... on this complex can bind to your biotinylated antibody. This creates a large meshwork of biotin-avidin...of some other common applications and links to relevant papers and webpages are listed below: EMSA (Electrophoretic...de, Rodriguez, P., Bonte, E., Krijgsveld, J., Katsantoni, E., Heck, A., … Strouboulis, J. “Efficient biotinylation...

...help save time by screening out irrelevant candidates, and, more importantly, it’ll help you define salary...contributed by Damien Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc. “How much...will take to secure your first (R-level) research grant.  Set your timeline from there, then start identifying...mouse experiments, start defining keywords you’ll want to find in candidates’ resumes or CVs; “vivarium...BEFORE searching for a faculty position. This is important to ensure that you’re looking at institutes that... try to keep conversations focused on research relevant objectives and their tangible resources. For example...Damien Wilpitz is a laboratory research management consultant and coach in Boston, MA. He specializes in assisting...

...run as The Green Lantern’s friend, St Walker: The Blue Lantern. While The Green Lanterns derive their power...Run4Rare team members complete the Boston Marathon, he wanted to run and play with my kids like any other youngster...Activin A. A commercially developed therapeutic antibody that binds Activin A and interferes with inappropriate...power from the emotion of will, The Blue Lanterns derive their power from the emotion HOPE; a fitting ...of rare disease sufferers like AJ. As the Blue Lantern Oath says: "In fearful day, in raging night, With...

...You've heard it before – it's important for scientists to get out there and talk about their work. To... is cool and why it really matters! There are important societal reasons, self-interested reasons, and...collaborative roles in academia, and that means it’s important to know how to talk about science to everyone ...myself) with a PhD in biology. Depending on what you want to do, that’s not necessarily bad news, but it is... understood why he cared to study mice on some distant island. He got a little money for his trouble, ...from the Ivory Tower.”  9)   If you think you might want to get serious about science writing, look into ...Communication Program at the University of California, Santa Cruz, has been turning scientists into confident...

...aligns with your interests. We've outlined some important factors to consider and summarize helpful advice...featured in "9 Tips to Achieve Success in Academia". Important factors to consider when choosing a research project... project After joining a lab, the next important decision to make is “What project will I work on?” In...like? What will be the impact on your field? You want to choose a research question that people (especially...critically evaluated your plan? Have you read the relevant literature to understand where the field is today...how can you mitigate them? If you were to write a grant for this project, what would be your critical experiments...typically require a diverse set of skills. It is important to know who in the lab and/or what facilities ...

...immobilized on a reduced glutathione matrix. The advantage of a fusion protein is the efficient purification...of an already immobilized target protein. The disadvantage is that the GST moiety, which forms dimers, ...binding kinetics of the target ligand. Another important consideration is whether the affinity of an experimental...removal of the fusion protein could provide a more relevant kinetic analysis, but the trick is to utilize ...receptor and subsequently evaluated interactions with mutant and wild-type erythropoietin molecules. The in ...We are clearly NEB “fanboys” as the bacteria significantly enhance protein folding (tested using real science... rabbit (EMD, AB3596) and mouse (NEB, E8032S) antibodies against MBP are commercially available. Factor...

...Attainable, Relevant, and Time-bound. It is a good idea to take larger goals (e.g. Figure out what I want to do... the helpful accountability that is one major advantage of working with a mentor.The responsibility of...mentoring relationship really productive, the most important step you can take is to set goals to work on together...to help a Mentee reach goals. One of the most important contributions made by a Mentor is to help the ...graduate student – you get the idea. There are many advantages to being a Mentor: Learn other perspectives ...reap rewards if you put in the effort. If you you want more great advice on mentoring, click below to download...

...of Different Methods for Generating iPSCs. One important note: in this table, reprogramming efficiencies... to tumor formation in iPSC-derived and iPSC-transplanted mice. Cre-deletable or Tet-inducible lentiviruses... be used to package reprogramming factors. An important milestone of reprogramming is the silencing of...cells for the presence of transgenes is still an important control. Additionally, Sendai virus doesn’t rely...express the reprogramming factors of interest. The advantages of this method are that it’s relatively simple...removed.  Replicating oriP/Epstein-Barr nuclear antigen-1 (EBNA1) based episomes: These plasmids carry ...substituting uridine with pseudouridine. The key advantage of using RNA to reprogram cells is that it’s simple...

...CRISPR/Cas9 could be used to edit several autosomal-dominant mutations that have been connected to the early...Chinese Academy of Sciences found that mice with a dominant mutation in a gene that causes cataracts could... Cas9 mRNA and a single-guide RNA targeting the mutant allele (7). An independent team led by Hans Clevers... chose to explore its potential in mice with a dominant cataract disorder caused by a single copy of a...of the cystic fibrosis report. “This is a big advantage over induced pluripotent stem cells, which need...and many others. “Those off-target effects are important,” she said. “We also really need to get a handle...optimization. In addition, the current work in developing anti-CRISPR proteins aid the limitation of these mutations...

... within tens of minutes (Cabantous et al., 2013). The Shu Lab took advantage of this and came up with ...signals are often difficult to detect, and thus quantify, due to small fluorescence changes of the donor...protease activity (To et al., 2016). However, the lab wanted to increase the fluorescence range to better study...on protease activation. Normally, β10-11 are antiparallel. To prevent binding, the β strands in FlipGFP...sequence, β10 and β11 can rearrange to their normal antiparallel position and fluoresce. FlipCherry: Red fluorescence...FlipGFP and FlipCherry at Addgene. References Cabantous, Stéphanie, et al. "A new protein-protein interaction...

...ideal for you. Here, we will chat about who might want to explore this role, how to seek out such an arrangement...them to a new institute and mentor that they may want to consider for future work placements. The chance...identified a gap or need in your research and decided you want to pursue a visiting position. What now? First, ... for the new placement. Make sure you read any relevant literature for the techniques you will be performing...these expenses, or you may need to seek out travel grants available from third parties or your home institute...generate data that will contribute to a publication? A grant? A patent? It is never too early to discuss authorship...feelers, apply to programs, and apply for travel grants. Good luck and happy travels! Resources Resources...

...studied for many decades, and they were known to be resistant to detergents and degradation under a variety ...different pH values. Using a screen that takes advantage of the fact that coiled R-bodies sediment in certain...see figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...initially used this screen to find R body mutants that couldn’t extend at a normal pH and later realized...Plasmids expressing expressing both wild type and mutant R bodies can be found at Addgene here. What can...F. R. Organization and expression of genetic determinants for synthesis and assembly of type 51 R bodies...trait: Absence of toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology...

...have gained or lost a specific gene survive (ex: antibiotic resistance marker). In a previous blog post, ... your plasmid backbone, as it often relies on antibiotic resistance. But what about identifying clones...𝛽-galactosidase activity. These cells, presumably the cell you want, cannot break down X-gal and are white. Cells that...to be strategic about your enzyme choices. You’ll want to pick enzymes that will give you different sized...to amplify anything. Like any experiment, you’ll want the proper positive and negative controls for your...provide all of the information you need, so you'll want to design a custom primer. Because Sanger sequencing...wouldn't be reached by sequencing from either terminus. Want to know more? Visit this blog post for tips for ...

...something like an antibody, enzyme, DNA, etc. The receptors job is to generate a quantitative signal upon recognizing...sense’ biological compounds and reactions; you can quantify them with biosensors! In this blog we will review...biological system. Basically, biosensors provide quantifiable readouts for levels of different chemicals, ...biosensor is the signal transducer which will quantitatively give a readout for whatever the bio-recognition...already engineered into viral vectors, so if you want to use them, you’ll need to use the vectors (or ...good thing, though! Viral delivery has several advantages over other mechanisms. For one, many viral platforms...

...site-specifically to alter its function, mimic a biologically relevant modification, or add in fluorescent or chemically... technologies have idiosyncrasies presents a significant barrier for those wishing to use it as a tool... or easily available, so you may find yourself wanting to use a protocol that you’ll have to rework to... They provide resources, knowledge, and most importantly, validated protocols (using readily available...their first protocol is available now. It is for recombinant expression of phosphoserine-including proteins... protocols come out! References Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C., & Schultz, P. G...Site-specific Incorporation of Phosphoserine into Recombinant Proteins in Escherichia coli. 12(21). https:/...