...Science: What the Future Holds and the Scientists Making It Happen, is an exercise in restraint. The very... revelations. The issues he discusses have been making the rounds in journal editorials, science social...likely range from mildly surprising to genuinely shocking. Mainstream media outlets have addressed CRISPR...recontextualized some areas of science that I’ve been thinking about a long time. For the scientists who like... Eric J. Perkins...

...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists...answered this question and used it to publish!  Working with AAV vectors in neuroscience Let’s consider...consider my first challenge when I started working with AAV vectors. I wanted to use an AAV vector to express...label neurons transduced by each serotype. Working at Addgene has shown me this was far from an isolated...Results from a survey conducted by Addgene in 2021 asking scientists what data would improve our AAV Data...

...PMCID: PMC110254. 3. In-vivo RGB marking and multicolour single-cell tracking in the adult brain. Gomez-Nicola...pre-clinical/early clinical stages. Researchers are also working to make lentiviral vectors safer. Since lentiviral...Winemiller A, Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl G, Tebas P, June CH,...therapy approaches to sickle cell disease. Hoban MD, Orkin SH, Bauer DE. Blood. 2016 Feb 18;127(7):839-48. ...

... a molecular leash for your protein. If you’re working with an uncharacterized protein, or a protein for...for antibody-based purification; has inherent enterokinase cleavage site GST Large Protein 26 Purification...context of a cell. FLAG tags can be cleaved by enterokinase, which recognizes DDDDK^X, cleaving after the...rounds of purification, with the cleavage reaction taking place between column runs. The TEV protease itself... Eric J. Perkins...

...Fujii: In the last several years, my lab has been working on development of technologies for biochemical ... molecules even enables us to skip the step of knocking-in. In 2013, we showed that isolation of a single...in the near future. Addgene: Has your lab been working on any other technologies that you would like to...that involvement in the scientific community is a kind of duty for researchers when considering the peer-review... in my lab then when they become independent. Looking for enChIP Plasmids? All Hodaka Fujii lab plasmids...

...that end, here are my personal 10 commandments for making it all work – most of the time. #1 Thou shalt find...happy at work, you will find balance. If you enjoy working and come home happy in the evening, your friends...of support to help with elder care, carpooling, cooking on nights when you can’t get home, babysitting ... really hard not to do this. For a family with working parent(s), the sooner your kids can make a decent...(except I am glad she never told me how much hitchhiking she did in Israel until after the fact). #9 Thou...

... used for protein localization, isolation, and tracking experiments (Rizzo, 2009). With many different...aggregation and cross-react with anti-GFP antibodies, making them difficult to use in a multicolor/multichannel...fluorophore-labeled antibodies with very specific reactivities making them more useful for multicolor imaging experiments...Davidson, and David W. Piston. "Fluorescent protein tracking and detection: fluorescent protein structure and...Blog Rabies and neuronal tracing Retrograde AAV: Making the journey from axon to nucleus AAVs CREATEd for...

...cargo Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker tape Teaching...effective and with no additional off-target edits, making them ideal for size-limited AAV delivery. PE6c ..., et al. (2023). Dynamic mapping of proteome trafficking within and between living cells by TransitID.... bioRxiv – doi: 10.1101/2023.02.07.527548.    Tracking cellular history with protein ticker tape By Brian...call for thin sections of tissue, the Wu Lab is working to create a three-dimensional atlas of protein ...

...amazing 111 countries (Figure 1)! But while we were making the materials available to many countries, import...stays in transit for a certain number of days and tracking average transit time for each courier we contracted... to increase accessibility to our repository by making it easier to find materials and learn about them...information are truly accessible, not just available.   Looking forward Over the past twenty years, Addgene has... accessible to scientists across the globe?” By asking this question, and by listening to the answers ...

...make sure your plasmid has a promoter suited to working in your host organism. Because transcription machinery... and there is very little crossover. Generally speaking, promoters in bacteria are less diverse and complex...elements, insulators, and silencers; however, some "leakiness" of transcription may occur. This is normally ...expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive  Widespread expression, but...optogenetics mRNA  Ca2+/calmodulin-dependent protein kinase II promoter Specific Used for neuronal/CNS expression...

...New to the lab? Or looking for ways to supplement your teaching? You’re in luck! Addgene’s Intro to the... We had many protocols for plasmid cloning and working with viral vectors, but what we lacked were videos...guiding completely new members of the lab or those taking introductory lab courses. The Intro to the Lab...science techniques. It’s also great for teachers looking to supplement their curriculum. From the details...

...the specific Rho GTPase/GEF pair (Ridley, 2016). Taking control of a RhoGEF is a great way to stimulate..., 2023). Rab family proteins support vesicle trafficking and organelle dynamics, but with dozens of family...Optogenetic inhibition of Rab protein vesicle trafficking via CRY2-CIB1 clustering. Image adapted from ...chemical composition of the membrane itself? The Baskin Lab previously published OptoPLD, an optogenetically....2015.08.005.  Tei, R., Bagde, S. R., Fromme, J. C., & Baskin, J. M. (2023). Activity-based directed evolution...

...express multiple integrins that bind the same ligand, making it difficult to dissect the function of one integrin...antibody is extremely rare. While many function-blocking integrin antibodies exist, almost all bind outside...Springer lab, reported the binding specificity, kinetics, and affinity of these 11 newly discovered synthetic...to the Springer lab, a highly specific function-blocking antibody, αVβ3.13, may also block function of ...important integrin on these cells. What’s next IPI is working with Addgene to accelerate biomedical research ...

...protein required for cytokinesis in JEI. Developing an interested in biology, working in a lab for the first...(the importance of word of mouth and personal networking!) and the manuscript review/submission process... first time, and thinking about the future. Many thanks to our guests,Olivia Ho-Shing, Mark Springel,...

...We’re making some moves over here at Addgene...and they’re looping? What’s the deal with looping...GIFs? We have answers! via GIPHY   We’ve been working hard to create the most informative and entertaining...protocol videos. via GIPHY   If you’re also looking for that perfect GIF for your TikTok or Instagram...us know on Twitter @addgene! Addgene is always looking for new ways to help you celebrate those wins in...

...If you’re looking for an antibody that works for your experiment, you may be overwhelmed by options —... — or not know where to start looking to find one. Luckily, there are websites that can help: Antibody...antibody catalogs at the same time.  If you're looking for an antibody, especially a validated antibody...antibody quality throughout research, consider checking out the Only Good Antibodies community on LinkedIn...

...Seeding Labs Plasmid Grant. For this grant, we’re working with fellow nonprofit, Seeding Labs, to distribute... health. 40:17 - 44:14 : Microbiome research - working with bacterial communities both on the human body...microbiome expertise. Many thanks to Kwabena Duedu for taking the time to sit down and talk with us.  Kwabena...the Addgene, Seeding Labs Plasmid grant and is working on a variety of projects in Ghana.     Special ...

...always gets done this time of year 😁 we stopped working on our reports/lab stuff for a few minutes to create...ladies #research #cardiovascularresearch #parkinsonsdisease A post shared by Ladies in labcoats (@ladiesinlabcoats...lab went all out to transform their lab into a walking tour of Buddy the Elf’s journey from the North ...fireplace scene.  Got our @EmoryChem lab office looking festive 🎄🌟🎁 @Addgene #DeckTheLab pic.twitter.com...

...material so that you can more easily find what you’re looking for. Whether you’re joining us for the first time...the blog that will give you a better experience. Looking for something? Use the navigation menu. The new... the top of the screen and type in what you’re looking for. After you search, you’ll see the blog articles...bloggers Like a blog post from a certain author and looking for more from them? Check out the new author pages...

...competent lentiviruses (RCL) especially if you are working on HIV positive human cell from the clinic. The...extra care has to be taken especially if you are working with proto-oncogenes or with human samples that... is now used as it shows rapid gene expression kinetics as compared to the Tet-off system. One drawback...one with either tTA or rtTA (FUW-M2rtTA). In this kind of approach, a population of cells might express...transgene resulting in low inducibility and potentially making it difficult to interpret your results. This can...

...when did you start thinking about them? 18:30 - 19:54 : How to think about picking a post-doc project ... you leverage your community, think hard about picking a good mentor, and begin considering career options...Mentoring" We'd like to thank Niroshi and Ben for taking the time to sit down and talk to us and wish them...

...a startup and back to academia. Chapter 1: How working at a startup prepared Tom for an academic career...more complicated biological devices. Chapter 6: - Making research go faster through Open Access. Chapter...investigate synthetic biology at Boston University, working in one of the founding groups of the field under... Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with Michael...

... including that it’s never too early to start networking! Learning about careers at Addgene Afterwards...popular, with the students excited to see a real working lab full of busy Addgenies preparing plasmid samples...impressed our guests, as did how much work is put into making sure scientists all over the world know that we...Advice from Our Bloggers Learn How to Improve Your Networking Skills  ...

...providing quality control and customer support, making plasmids easy for you to find and request in our... Addgene? Here's an inside peak into the daily workings in Addgene's lab. We ship individual plasmids ...Stabs are organized by shipment and moved to our packing area.Using the barcode system, the correct plasmids...necessitate an office move. We've got ~50 Addgenies working to ship plasmids, analyze QC sequence, answer scientists...

...Brownstein, an Addgene board member. When I was working in Human Resources at Harvard Medical School, Joanna...Joanna was my colleague. She mentioned Addgene was looking for someone with HR expertise to join their newly... her forever home. I feel blessed that I’m not working so that I’m available to spend time caring for ...for my mother. For fun, I enjoy pilates, jewelry making, reading, sailing, and gardening. Learn more about...

...expression cassettes (Shaw et al., 2023). Those looking to express high levels of their gene of interest...more! Engineering disease resistance through gene stacking, increasing production of a protein of interest...requirements and editing abilities, such as base editing, nicking, cleaving, etc. The possibilities really are endless...mammalian systems and Drosophila. Similarly, the Multi-Kingdom (MK) Golden Gate system was designed to work in....2024.100519. Chiasson, David, et al. “A Unified Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports...

... don’t want to spend months doing surgeries and making fiber optic cables only to hook up your lasers ...alter the light spectrum for activation and the kinetics of opsin activity/inactivity. Dr. Deisseroth has...(not expressing the opsin) are also available. Picking your opsin  You must consider whether you wish ...neuronal firing, the cell-type specificity, the kinetics of the opsin variant, and other particularities...the promoter of the cre recombinase while still taking advantage of the robust expression via the promoter...

...Francisco International Airport listening to the Lion King soundtrack and writing this post, it is my pleasure...what new post contributed the most to this record breaking month and to find other posts that deserve a second...Navigate a Scientific Enterprise in Flux CRISPR Kinome Libraries Available: Pooled and Individual Plasmid... Research Community Grad School Advice Part 1: Picking a Lab and a Project That’s it for now. As I indicated...

...Tools for Making Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for making scientific...we're gearing up to start a new decade, we’re also looking back at the last year of scientific discovery. ...Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by Joachim Goedhart Guest...

...at a lindy hop swing dance event,” Zheng says (networking can happen anywhere!). The pair quickly teamed... in November 2017. #Phage researchers! I am working with a team to get Burkholderia cepacia phages ...must recognize a specific part on the bacterium making each phage is very specific. Some infect only one...about scientific sharing Find blog posts about networking Additional resources on Addgene.org Find molecular...

...excited to finally share with you what we’ve been working on behind the scenes here at Addgene.  Addgene's...messaging, as well as simplifying the page, and making it all look clean and easy to use.  Easily search...global navigation and menus over the past year, making it much easier to get around our site.  Community...smile, but occasionally you might notice Blugene looking sad, especially if an experiment doesn’t go as ...

...experiment. (And you’re certainly not alone – we’re all looking for high-quality antibodies and reagents that work...and sorting. Reference images can be viewed by clicking on the links in the "Agree" and "Disagree" columns...associated metadata adapted from the Affinity Reagent Working Group of the Human Biomolecular Atlas Program (...community when designing the IBEX Knowledge-Base, seeking to create an iterative process that is constantly...

... search. This is especially true of scientists seeking a first job after the academic bench. Utilizing... the job posting instructs even if you have a networking connection that will be helping (see below). ...strengths in your resume and don't be shy about asking for help. The worst thing that can happen is someone... scientist networking...

...,” said Addgene’s Lianna Swanson, who has been working with members of Davidson's lab to coordinate the...For more information on choosing a backbone and making fusions, see this blog post from Joachim Goedhart...Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection.  mEmerald plasmids and many... many more! Baird has a helpful tip for anyone looking to peruse the collection without getting overwhelmed...

...paper they’re reading has been linked in a ReFigure making it easy for others to discover your insights on...researcher Christa Osuna collated Zika case studies, making a meta analysis available to the community on ReFigure...PubMed Central, eLife, PLOS and Figshare but we are working on expanding our reach. We can’t save you the hard...career researcher and cofounded ReFigure with Joe Akin. She envisions a world where the impact of teaching...

...faster kinetics. The opsin ChETA, for example, is a ChR2 opsin mutated to have faster off kinetics and allow...set of tools therefore depends on what you are looking to study. Figure 1: A schematic showing the...receptors in the brain. One way to address this is by working with organisms with light accessible brains, such...

...and you are smart, you can be a good manager. The kind of smarts and the type of skills that it takes to... from being included in organizational decision-making. It is imperative that employees are thanked for...manager keeps these factors in mind at all times when making decisions, assignments and polices. Sometimes the...priorities for advancement in science Have their own working schedules and may not necessarily be comfortable...

...persuing milestones and working hard. Being patient, being selective, and talking to peers and mentors will...with the current technology to identify a problem, asking what approach will I take and what can I learn,... Dr. George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists...

...immunohistochemistry images that are crisp, bright, and lacking in non-specific staining can be a challenge in ... mixing your primary and secondary antibodies, blocking excess secondary with serum from the same species...problem.  Pre-block with Fab fragments (Figure 2C) Speaking of Fab fragments… Another strategy for species-on-species...0022155413511620  Lu QL, Partridge TA (1998). A New Blocking Method for Application of Murine Monoclonal Antibody...

...just on the disease front. Viruses survive by hijacking the machinery of the cells they infect. Once they...proteins. This is, for the intrepid researcher looking for molecular biology tools, a very useful trait...Created using biorender.com   If you’re new to talking about viruses, here’s a quick tip on terminology...reproduce only in experimentally controlled conditions, making them far safer to work with than virions. The four...

...whirlwind of skill acquisition. Classes covered filmmaking, podcasting, philosophy of science, creative ...life of Srinivasa Ramanujan, to be precise) and remarking to a classmate that, if this was one day required...communication is an equally fruitful endeavor for those embarking on the route towards academia. Many of those entering...master's degree in 2020, his research explores risk-taking behavior among collision sport athletes, as well...

...Working on the scientific support team at Addgene, I get to see everything that can go wrong with a plasmid...DNA yield. Test multiple, small colonies After streaking LB agar plates, you may notice that there are ...recombined product tend to grow faster, so we recommend picking the smaller colonies which are more likely to carry...the recombined vector backbone, you can try re-streaking for single colonies and testing multiple preps...

...features provide extra layers of risk management when working with viral vectors. Typically, one day after transfection...particles are secreted into the cellular media, making viral collection fairly easy. Producer cell media...transfect’ cells lines are amenable to transduction, making it a logical choice for efficiency. Where viruses...with alternative methods. Ready, set, transduce! Making virus can sound daunting, but it is easier than...

...inclination to be a teacher was looking for an outlet. I started asking people: why do you do what you ...bioscience into context, while at the same time working through the details of experimental design and ...of science communication professionals who are thinking about, for example, how to use new technologies...

...Plasmagenes? In the 1940s and 50s, scientists were working to understand genetic cytoplasmic factors that ...and we’ve been using "plasmid" ever since! From napkins to notebooks Although discovered in the early 1950s...initiated at a Hawaiian deli in 1972. Using a deli napkin for paper, a small group of scientists including... curate, and distribute them all in the name of making it a little bit easier for scientists to conduct...

...Will the job require public speaking? Will you be leading a group or working as part of team? Will you .... It is all too easy to get tunnel vision when working towards a specific goal, and when you realise that...often be misleading. Key aspects to determine are working hours (9am-5pm Mon-Fri or overtime and weekends...

...established in yeast, including a widely used Parkinson’s experimental model. In fact, approximately 25%...phaffii is highly efficient at protein secretion, making harvesting recombinant proteins a cinch. This is...it doesn’t secrete its own endogenous proteins, making the recombinant protein its major secretion. Beyond...players in the game (even beyond K. phaffii ) worth looking into whether you’re just dipping your toes into...

...amplification looks like, but by looking at these kinds of graphs, making sure few/no plasmids have dropped... the key points you’ll need to remember when undertaking library amplification and sequencing. Maintain...

...will define positive selection as the means of picking up positive traits (i.e. the plasmid contains a...toxin), whereas negative selection would be the picking up of a negative trait (i.e. the plasmid contains...Conferring Resistance Cell Types* Mode of Action** Working Concentation*** Blasticidin bsd  HeLa, NIH3T3...transfection when creating stable cell lines. Looking for Mammalian Expression Plasmids? Check Out Addgene's...

...spread of COVID-19. That’s why we’re proud to be working with Ginkgo Bioworks to synthesize and share over...analyzed first thing the following day. Nothing like waking up to fresh data! These same plasmids are then ...ensuring our rigorous QC standards are met, we are taking a unique hybrid approach compared to our standard...high throughput platforms, Addgene and Ginkgo are striking a balance by modifying certain processes in both...