...new to Addgene this year. Feel the love with MTRIA sensors One of the latest additions to Addgene’s catalog...imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator using small...on the Addgene blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent...Here at Addgene’s headquarters, the skies are getting dark and wintry with the end of year approaching...other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted GFP into an...localized for synaptic glutamate response. They generated variants optimized for different applications ...St-Pierre lab and colleagues have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-...

...learning.” Addgene’s Director of Product Management Eric Perkins, who has been in many roles in Addgene since...work at Addgene. I posed this question to all nine of my interviewees. Fabienne Furt, Addgene’s Associate...resources on the Addgene blog Browse our science career blog posts Download Addgene's science career guide... guide Resources on Addgene.org Learn more about working at Addgene ...Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst intern. As a member...go back to school right away. Shannon Rinaldi, Addgene’s Lab Manager, was taking classes at her local community...community college when she took her first job at Addgene as an Order Processing Assistant. After a few months...

...Resources on the Addgene Blog Learn About the Addgene Help Center Get Help with the Addgene Deposit Process...emails in my six years as a senior scientist at Addgene. Having spent that long in customer service, I'...Though my experience is based solely on my time at Addgene, I’m confident that these tips and tricks will ...another great source of information. At least at Addgene, the questions in our FAQ section are really, truly...not realize it, all of the reagents available at Addgene come from researchers, and the vast majority of...that for you” meme is infamous enough to have generated it’s own tongue-in-cheek website.  If you are ...happens more frequently than you might think. The Addgene scientists answering the phone are smart, but they...

...from addgene.org  Plasmids encoding nanobodies at Addgene Sybody Generation Toolbox from Addgene     ...only antibodies. (A) Overview of genetic modifications made to generate the nanomouse. (VHH = variable ...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended...to human populations around the world. Here at Addgene, we love camelids too, though not just for the ...chromobodies,” which are single-domain antibodies genetically fused to fluorescent proteins and used to visualize... synthetic single-domain antibody libraries to generate new sdAbs. There has also been some interesting...) (Xu, et al., 2021).      Figure 2: Genetically modified mice offer an alternative method to ...

...been generated for yeast (Kim et al., 2019) and fruit flies (Bischof et al., 2018), the generation of such...quantities (and the correct stoichiometric ratios) to generate useable data. What’s more, measuring and quantifying...straightforward nature of BiFC also makes it easier to generate a large number of constructs for high-throughput...screening of PPIs. For example, Bischof et al. (2018) generated an open reading frame (ORF) library to screen ... such resources in less genetically tractable organisms lags behind. BiFC can also be used to screen for...pair. Find BiFC plasmids for your experiment at Addgene! How to Design a BiFC Experiment Much like FRET...optimized to be successful. A BiFC experiment generally involves three steps: fusion protein construction...

...scientists are generous Besides some of the scientists I've already mentioned, Addgene depositors Marius...What an honor for Addgene to be a part of that!  Additional Resources on the Addgene Blog Learn about...landmark Cell paper showing that it was possible to generate induced pluripotent stem cells (iPSCs) capable...called OSKM factors). As the Yamanaka lab's primary Addgene contact for many years, I've been looking forward...example of the fast movement in this field.  As an Addgene scientist, I'm exposed to many scientific communities...clinical trial patients with advanced macular degeneration; Laurenz Studer discussed his work with Parkinson's...quality control process alone takes nearly a year. Addgene depositor Juan Carlos Izpsua Belmonte said it took...

...communication blog posts Resources on Addgene.org Find careers at Addgene ... project I’d seen in my already long tenure at Addgene. I would work with our research team and software...pinpointed and (mostly) solved two main problems with Addgene’s cross team communication. Although these real ...these plates now” to a fellow team member. But, at Addgene, we use seven different types of “plates” (Fig....not the container was barcoded (this is pretty Addgene specific) The types or purposes of each given container...that there were a lot of words that nearly all Addgene teams used that seemed simple, but they could in...solution for this problem was the creation of an Addgene lexicon. The base lexicon was and continues to ...

...approaches to control transgene expression by restricting expression to genetically defined cell types. ...neuroscientists have begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases...vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation of specific neuronal...changes affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed...molecular tools and sensors Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV.... Specific transgene expression can be also achieved in nontransgenic animals by using cis regulatory ... one location can result in expression of an optogenetic payload in neurons that project to different ...

...CRISPR, Addgene gave to me: a primer on prime editing.  On the eleventh day of CRISPR, Addgene gave to...all of them here.  On the first day of CRISPR, Addgene gave to me: a miniature CRISPR-Cas system for mammalian... mammalian use. On the second day of CRISPR, Addgene gave to me: engineered pegRNAs to improve prime editing...editing efficiency. On the third day of CRISPR, Addgene gave to me: Selective Endogenous eNcapsidation ... Delivery (SEND). On the fourth day of CRISPR, Addgene gave to me: enhanced prime editing systems through...manipulating MMR machinery. On the fifth day of CRISPR, Addgene gave to me: efficient CRISPR editing with a hypercompact...engineered guide RNAs. On the sixth day of CRISPR, Addgene gave to me: PRIME-del (Precise genomic deletions...

...analyze and annotate any DNA sequence. SnapGene Viewer: The free SnapGene Viewer is great for looking at plasmid... biology tools.  Addgene Data Hub: A data repository for validation data on Addgene/CLOVER viral vectors...Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA.   DNA / .../ protein database search and analysis tools Addgene's analyze sequence tool: Annotates features, BLASTs...restriction enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled...available sources, as well as those deposited with Addgene. NCBI BLAST: Compares nucleotide or protein sequences...tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence manipulation...

...preps available in the Addgene viral vector repository! This includes the first Addgene prep packaged in the... AAV1 Optogenetics Karl Deisseroth New serotype pAAV-hSyn-hChR2(H134R)-EYFP AAV2 Optogenetics Karl Deisseroth... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...other new AAV vectors as well — including next-generation biosensors for GABA, dopamine, serotonin, and...Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor...serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV5 Optogenetics Ofer Yizhar New viral prep pAAV_hSyn-DIO-PdCO-mScarlet-WPRE...-PdCO-mScarlet-WPRE AAV1, AAV5 Optogenetics Ofer Yizhar New viral prep with multiple serotypes pAAV-Syn-FLEX-rc...

...C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first anti-c-Myc [9E10] (...high-throughput screening platform to apply saturating mutagenesis on 21 positions in the protein sequence of the...approach and outcomes of the screen for improved genetically-encoded voltage indicators (GEVIs) based on the...kinetics. They demonstrate the use of HcKCR1 as an optogenetic silencer in experiments with mouse cortical neurons... HcKCR2, making it more suitable for in vivo optogenetic use.   Fig. 2: A) HcKCR1 is highly selective...control K+ gradients and can be used as a potent optogenetic silencer. The current work also proposes promising... light-gated potassium channels that mediate optogenetic inhibition. Nat Neurosci 25, 967–974 (2022). ...

...pAAV-hsyn-flex-Voltron-ST DREADDs / Chemogenetics AAV We have also expanded our inventory of chemogenetic AAV for activating...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...jGCaMP7c for calcium detection PSAM4 GlyR for chemogenetic inhibition Flp expression for intersectional...FDA-approved drugs. See our inventory of DREADDs / Chemogenetics AAVs New DREADDs Plasmid Serotype Name ...AAV5 pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) New chemogenetics AAV Plasmid Serotype Name 119741  AAV5 ...pEF1a-DIO-FLPo-WPRE-hGHp Other new viral vectors New optogenetics inhibitors Plasmid Serotype Name 99039 ...These vectors listed below should be released at Addgene in the next 6 months, pending quality control. ...

... Here’s how Addgene helps: For Providers: One-time Deposit: Simply deposit with Addgene (it’s FREE!)...Streamlined Processing: When you place an order, Addgene generates the MTA and emails it straight to you, or ... on collaboration (it’s the heart and soul of Addgene’s mission), but legal complexities can arise when...institutions require them for material transfers. When Addgene entered the plasmid sharing space, we heard over...confidence. Benefits of the UBMTA: Peace of mind Addgene has adopted the terms of the Uniform Biological... review by the Provider Limited liability Addgene makes it easy Even with all those benefits, we ... handle all the paperwork. Record Keeping: Addgene will let you know exactly how many times your materials...

...protocols Sep-OTSλ plasmid - Addgene # 68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG...E17TAG GFP plasmid - Addgene #68295 SupD plasmid – Addgene # 68307 For further technical reference and protocols...candidate regulatory kinase is known, it’s possible to generate non-phosphorylated recombinant protein and perform...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...flavors’ of Sep-OTS plasmids are available through Addgene, but we have worked most extensively worked with...must be recoded to ‘TGA’ using site-directed mutagenesis. In the C321.ΔA strain, this TGA will be recognized...position. Practical tips for your experiments To generate as much pSer-incorporated protein of interest ...

... biology over the past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers...proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview by Erik Snapp Addgene depositor...2020! Additional resources on the Addgene Blog Read other Addgene news Find the most recent plasmid ...posts Resources at Addgene Browse the repository Find educational resources at Addgene Search for plasmids...discovery. We’ve documented many new tools on the Addgene blog and have created and updated guides on various...blogger Joachim Goedhart shares some strategies for generating GFP fusion proteins. From considering protein... by leaving a comment or emailing us at blog@addgene.org.  Thank you again for a fantastic year and happy...

...executive director of Addgene, it is my privilege to uphold the values that shape Addgene’s mission and work...thrilled to be sharing our plans for 2024.  In 2023, Addgene distributed 188,729 materials to 39,844 scientists... three–year project that touched every team at Addgene. As a resource, it will allow scientists around...Institute for Protein Innovation (IPI), in which Addgene will distribute recombinant antibodies (rAbs) developed...focus on expanding international access to the Addgene repository, partially through work with our distributors...mention that 2024 is an incredibly special year for Addgene: it marks our 20th anniversary. Throughout the ...months, we’ll also share our work in evolving Addgene’s strategic vision for the future. Over the years...

...technique, you first perform a round of PCR to generate DNA fragments with 15-40 bp of homology at their... site, toposisomerase I, and a taq polymerase-generated insert. When mixed, topoisomerase I provides both...Taq perfectly anneal with the topoisomerase I generated overhangs in the backbone. Once combined, the ...processes will require a PCR reaction or two to generate templates for the cloning process. Time spent ...spent actually doing the experiments are also generally comparable so the best way to compare costs is to ...assembly is a highly versatile and efficient way to generate a wide variety of constructs. Similar, Gateway...$20-30 per reaction but can be used to quickly generate multiple clones. Difficulty: If you’re an experienced...

.../nn.4265.  Additional resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit... al., 2024). They deposited their plasmid with Addgene, hoping to inspire others to explore the phenomenon...Although a wide range of fluorescent proteins and optogenetic tools exist, humans (like many organisms) are...magnetoresponsive fluorescent protein. Controlling an optogenetic or fluorescent tool with magnets would enable... molecules like proteins with magnetic fields generally appears to be weak — if they exist at all. Andrew... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...

...perfect job.   Resources at Addgene Read Additional Career Posts on the Addgene Blog Browse Our Careers ...a full time position in a lab. I worked on the genetics of Wilms Tumour, which is a type of pediatric ...study a Master of Science (MSc) in Human Molecular Genetics to give me an edge over other graduates during...costs. My MSc courses were interesting and luckily genetics is a sector which has been rapidly growing. These...factors left me optimistic that I would find a genetics related role soon after finishing my MSc. The ...Australia, this was a perfect opportunity to work in a genetics lab. As a UK citizen, I was entitled to a Work-Travel...the best outcome for a specific combination of genetic mutations. This is a particular challenge in pancreatic...