...If you’re been following Addgene on #souptwitter or if you just enjoy a warm meal on a winter day, you...molecule into the cell. The fluorescence that is generated allows for qualitative and quantitative measurement...placing the aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the... we’ll take a deeper dive into the content of Addgene’s aptamer soup by exploring the development and ...and applications of fluorescent RNA aptamers in Addgene’s collection. Green fluorescent aptamers Spinach...s group created Spinach2 using site-directed mutagenesis of Spinach. In doing so, they identified the ...changes in intracellular viral rna." Journal of General Virology 98.5 (2017): 992-1003. PubMed PMID: 28555544...

...Resources on Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene Viral...Viral Services Resources on the Addgene blog Viral Production at Addgene Viral Vectors 101: What is a Virus...infiltration into a cell. The twist is that the typical genetic cargo — the viral genome — is swapped, at least... viral genome has been swapped for alternative genetic cargo, they can’t reproduce more functional viruses...   Is there an easier way? In fact, there is! Addgene has a number of ready-to-use viral preps available...

... autophagy biosensors currently available from Addgene. Rosella autophagy biosensor Rosella is a dual-...-targeted and mitochondria-targeted Rosella at Addgene! Keima autophagy biosensor Keima is a mammalian...Find the mitochondria-targeted Keima variant at Addgene! mRFP-GFP-LC3 tandem fluorescent protein This autophagy... red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3∆G: A fluorescent probe that releases...translation, this fusion protein is cleaved to generate one molecule of GFP-tagged LC3 for every one molecule... flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References Klionsky, Daniel J., Ana Maria Cuervo...PMCID: PMC5618514. Additional resources on the Addgene blog Beware of fluorescent protein oligomerization...

...his genetically encoded magnesium sensor, the first of its kind. “I hope by depositing at Addgene, people...Institute for Neurobiology has been working on genetically encoded indicators of calcium and other small...Twitch calcium sensors, which are available at Addgene, can be used for ratiometric in vivo imaging. Griesbeck... of the standard organisms for which extensive genetic tools are available. While he recognizes that some...PubMed) Lindenburg et al. MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. ...(PubMed) Interested in More Hot Plasmids from Addgene? Check out our Hot Plasmid page, where we highlight...some of the popular & newly available plasmids at Addgene. If you're interested in plasmids for in vivo imaging...

...began working remotely recently? Most of us at Addgene began working from home last week to flatten the...new. We usually have yoga class twice a week at Addgene HQ, but now we’re sharing links to online yoga ...before bed will help you prepare for sleep. At Addgene, our mindfulness group is sticking with their Headspace...virtually connected with the people in your life. At Addgene, we’ve started virtual activities like team lunches...being safe. @GHP_HarvardChan — Shekhar Saxena (@ssaxenageneva) March 16, 2020 Give back to your community...societies hosting virtual networking events. Download Addgene’s Science Career guide to learn more about career...framework to design your future experiments.  Use Addgene’s AAV Data Hub. You can search the AAV Data Hub ...

... pAAV.hSynap.iGABASnFR (112159-AAV1) Optogenetics Optogenetics continues to be a useful method for manipulating...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...serotype testing Calcium sensors and GABA sensors Optogenetics Recombinases And more! Control AAV For many...Recombinase AAV Targeting and manipulating specific, genetically-defined neuronal populations with intersectional...pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE (104496-AAVrg) Optogenetics pAAV-hSyn Con/Fon hChR2(H134R)-EYFP (55645-...55645-AAV5)  Additional resources on the Addgene blog Read all of our viral vectors blog posts Find tips...

...more. Being that this is the Addgene Blog, I’ll, of course, mention Addgene as a source of sequenced and...event, I gave my own talk on reproducibility at Addgene and here I summarize what I learned both in preparation...31 - 49:45 - Reagent Sharing - Susanna Bachle, Addgene the nonprofit plasmid repository 49:45 0 1:18:45...new ways of understanding how the body works, generating new therapeutics, and sometimes formulating new...viral vectors. When working with a plasmid from Addgene, you can be confident that you’re using the actual...hosted by individuals and by organizations like Addgene (a good example is our series of blog posts on ...Initial positive results and the excitement they generate can, however, bias us. We’ve all heard stories...

... on the Addgene blog An Overview of AAV Encoded Calcium Biosensors Illuminating Epigenetics with a ... systems that measure biological phenomena by generating an output which is directly proportionate to ...antibody, enzyme, DNA, etc. The receptors job is to generate a quantitative signal upon recognizing the analyte... (fluorescence, pH, etc.). The transducer will generate a numerical value which represents the amount ...: Almost all of the biosensors discussed are genetically encoded, and luckily, we have pre-designed plasmids...disciplines! Most of the sensors designed for calcium generate a fluorescent output, which will turn on, change.... DOI: 10.3389/fncel.2019.00474 Resources on Addgene.org Fluorescent Proteins: Biosensors Viral Service...

...and Jennifer Doudna Labs used targeted mutagenesis to generate hyper accurate Cas9 (HypaCas9). This nuclease...frequently used Cas9, spCas9 from Streptococcus pyogenes, this sequence is 3’-NGG. But Cas9s found in other...exhibit enhanced site accessibility In order to generate a genomic alteration at a specific location, it... sites. Using a combined random and directed mutagenesis approach, the Anna Cereseto Lab created pX-evoCas9...the Jungjoon Lee Lab used directed evolution to generate Sniper-Cas9 resulting in a Cas9 with improved ...PMCID: PMC6255809.  Additional resources on the Addgene blog Read our CRISPR 101 blog posts Find more ... Download the CRISPR 101 eBook Resources on Addgene.org Visit our CRISPR plasmids and resources page...

...and learn more about Addgene’s viral service. Additional resources on the Addgene blog Download our Viral...service, and now we’re “going viral” again! #AddgeneGoesViral on Twitter Starting today, our Twitter feed... in the graphic below. Follow the hashtag #AddgeneGoesViral to find information on the basics of viral...applications, and some new resources available from Addgene. There’s a lot to talk about! And by the way, there...reagents and data, we’re also publishing research. Addgene’s research team also investigates new strategies... posts Learn more about AAVs Resources on the Addgene website Visit the AAV Data Hub Learn more about...

...developed a dual virus system using both optogenetics and chemogenetics to study the activity of parvalbumin...in the lab. He has recently developed a third-generation intensiometric glutamate sensing fluorescent ...catalyzes the desulfuration of 3-mercaptopyruvate and generates an enzyme-bound hydropersulfide that is then transferred...withdrawal before they occur.  First, she used a genetic method to tag neurons activated by opioid withdrawal...light activates the neurons (via oChIEF) and a chemogenetic agonist inhibits the neuron (via hM4D). She ...postdoc at Harvard University where she engineers genetically encoded voltage indicators. Tian developed a ...optimize a red‑shifted archaerhodopsin‑derived genetically encoded voltage indicator which had improved ...

...are tumor microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression...that can be transferred to other cells, inducing epigenetic changes in targeted cells. In this case, exosomes...formation by inhibiting cell growth, invasion, and angiogenesis (Allinen et al., 2004). In fact, Sanford H. ...influence tumor microenvironments: immunology and angiogenesis (Witz et al., 2009). Then, scientists began ...Fernando Schmitt. "Microenvironment in breast tumorigenesis: Friend or foe?." Histology and histopathology...PubMed PMID: 20036813. Additional resources on the Addgene blog Read our cancer related blog posts Learn ...about the tumor microenvironment Resources on Addgene.org Visit our cancer research plasmids and resources...

...mouse inner ear. Fourth-generation cytosine base editors The fourth-generation base editors, BE4, reduce...activity.  Find CRISPR base editing plasmids at Addgene Improving cytosine base editing scope and efficiency...18 bases upstream of the PAM.  David Liu’s lab generated BE3 variants with other deaminases: AID, CDA1,...these three improvements represent the fourth generation of base editors. Compared to BE3, BE4 offers ... phage-assisted evolution selection systems to generate ABE8e(TadA-8e V106W), which edits ~590-fold faster...., 2020). This is important because ABEs have generally been slow, meaning that the Cas9 domain often ... N. M. (2023). Improved cytosine base editors generated from TadA variants. Nature Biotechnology, 41(5...

...Novartis, Biogen, Amgen, New England Biolabs, Regeneron, and Genentech, among others, and the list is growing...towards researching neurodegenerative diseases.   Additional resources on the Addgene blog Check out all...choosing a lab Resources on Addgene.org Find open positions at Addgene Check out our career videos ...beginning of March, and the jobs are mixed into the general careers section of their web site. Each position... a position open up. Other companies, such as Genentech, will not. The point is that each company has ...

...488nm blue light. The T203H mutation in PA-GFP generates a chromophore population that is more neutral,...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...negative” photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't...endogenous protein structure and function. By generating monomeric PA-FPs, there is less chance that the...induce though their continuous excitation. By generating PA-FPs fused to intracellular proteins, scientists...PubMed PMID:18724362. 8.Stiel, Andre C., et al. "Generation of monomeric reversibly switchable red fluorescent...PMC4344411. 13.Berlin, Shai, et al. "Photoactivatable genetically encoded calcium indicators for targeted neuronal...

... use.  View the Addgene Plasmid Request Data Dashboard! Figure 1: The Addgene Requests Dashboard...I was excited to see that Addgene has recently added a new feature for depositing labs to its website...default. Depositors can download their CSV file from Addgene by following these instructions, and then uploading... 2: Insert showing the upload function for the Addgene Requests Dashboard After the upload, the name...displays the data in a standardized way. Since Addgene encourages the public sharing of aggregated data...Amsterdam). He develops, characterizes, and uses genetically encoded fluorescent probes. You can follow him...

...Antibodies Resources on Addgene.org Find recombinant antibodies at Addgene Find plasmids encoding recombinant...introduction, we broke down the method into six general steps and outlined the considerations to be made...fixation*. Organic Solvents Organic solvents, generally acetone, methanol, or ethanol, work by dehydrating...to use your specific antibody for IF, look for general IF protocols for your sample type. Starting with...cross-linking fixation; they require an additional step generally referred to as Antigen Retrieval. I chose not ...the-process-of-fixation-and-the-nature-of-fixatives/  Additional resources on the Addgene blog: Antibodies 101: Introduction to Immunofluorescence...recombinant antibodies at Addgene  ...

...science communication Resources on Addgene.org Learn how Addgene engages with the community ... Addictive Brain and science communication for general audiences. How does The Addictive Brain address...findings, and experiments as articles written for general audiences. So far, we are beginning to make an ...science, especially disease related topics. Neurodegenerative and brain disorders are frequently overshadowed...“manipulate” could mean “illicit tampering” to general audiences while “manipulate” could mean “worked...BioTechniques.     Additional resources on the Addgene blog Find all of our science communication posts...

... The HuidaGene Therapeutics team has deposited their editors, called gTBE and gCBE, with Addgene. gTBE...editors were developed, though, researchers at HuidaGene Therapeutics guessed that fusing MPG to an ABE...this promising start, the team used rational mutagenesis to produce the adenine transversion editor AYBEv3...and ABEs, they could be useful for saturation mutagenesis, and they provide a strong proof of concept for...empty space. Over several rounds of rational mutagenesis, the team progressed to gGBEv6.3, which was capable...s41467-024-50012-w Additional Resources on the Addgene Blog CRISPR 101: Cytosine and Adenine Base Editors...about four base editor reporters Resources on Addgene.org Browse CRISPR plasmids for base editing Get ...

...Resources on Addgene.org Find protocols of molecular biology techniques Find plasmids at Addgene Make your...have integrations with a broad range of relevant generic and science-specific tools. In addition, it should...repositories Over the past decade or so a number of general purpose repositories have been launched and are... associated with publications into one of five general purpose open source data repositories: Dataverse...one and preferably more than one of these five general purpose repositories. RSpace has been integrated...@rory_macneil.     Additional resources on the Addgene blog Find tips on choosing an electronic lab notebook...your work more accessible by depositing at Addgene ...