...adenine base editor Earlier this year, Keith Joung’s lab developed SECURE cytosine base editors, which decrease... Nature Biotechnology Find the base editors   Single base editor that induces C-to-T and A-to-G mutations...developed a base editor that combines the abilities of C-to-T and A-to-G base editors into one protein...pyogenes Cas9. They called this editor Target-ACE (adenine and cytosine editor). Target-ACE exhibits increased...research! This time: CRISPRi in Burkholderia Base editors CRISPR toolkit for epitope tagging Lentiviral capsid-based...APOBEC1. Now, they’ve engineered SECURE adenine base editor variants using a protein-truncation strategy with...a target site compared to a mixture of two base editor enzymes in mammalian cells. Read the paper in ...

...activation of keratinocytes, which release inflammatory mediators that trigger the activation of dendritic...recruitment of neutrophils. Though there are IL-36 receptor inhibitors in development, there’s still a lot of mysteries...Generalized pustular psoriasis is a chronic autoinflammatory skin disease estimated to affect 0.27 to 4.6...actually get out of keratinocytes and induce their effector functions. It’s an important process that we need...IL-36 cytokines are a potent inducer of an autoinflammatory loop: keratinocytes both respond to and release...when he was studying thoracic aortic disease. “My mentors Dr. Dianna Milewicz, Dr. Carolyn Bondy, and Dr....with TS (Trolle et al., 2016). Using lentiviral vectors from Addgene, the lab will express these candidate...

...point.   What makes a story? A story is a narrative connecting cause and effect. Stories can and should be... or notes. Listen to their stories and narratives (see below “Tell a story”) and note how they build suspense...hear material delivered in the form of a story. Using stories is one of the best ways to make you and ...me and wished I had been more careful. 3. Tell a story We are biologically programmed to listen and remember...presentations engaging and memorable. If you read my story above about the presentation faux pas, you are more... striking images, creative ideas, and engaging stories as you encounter or experience them. Drop content... all graduate students and postdocs form peer mentoring groups to practice giving talks and getting feedback...

...lab has developed AAV vectors to selectively visualize excitatory and inhibitory synapses in the brain...combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory synapses... find in this post: Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein system...New from the viral service Labeling excitatory and inhibitory synaptic proteins with FingRs By Jason... proteins PSD95 and gephyrin to label excitatory and inhibitory synapses, respectively. Once expressed...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles... cycle in vivo, particularly if the cells are migratory or in deep tissues. To resolve this issue, Shangqin...

...editing assemblies are sensor-actuators, either transcriptional regulators or chromatin-modifying enzymes...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...s chromatin status into account (Fig 1a). The laboratory of Karmella Haynes at Emory recently contributed...contributed a set of plasmids to the repository that will help epigenetic researchers custom design their own...Examples of histone-modifying enzymes and reader-effector proteins. b) Golden Gate assembly (backbones available...with examples of potential synthetic chromatin mediators and probes. Image from Priode et al., 2021. ...potential tools fall into two categories: probes and editors. A probe array may consist of at least two sensor...

...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). ...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...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...

... 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 ...

...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...

...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...

...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...

... 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...