...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...

.../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...

...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...

...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...

...s41587-023-01833-5 Resources on Addgene.org Optogenetics guide Chemogenetics guide BRAIN initiative plasmid... on the Addgene blog Plasmid tools for microbiome studies Channel rhodopsins for optogenetics Viral Vectors...electric stimulation, drug administration, and optogenetics. Enter neural fibers! The neural fiber tool...features enable the fibers to emit light for optogenetic purposes, measure temperature, study electrophysiology...molecules.   Fun fact: A 1 cm block of polymer can generate many meters of fibers!     Fig. 1 - ...the labs looked at previous implementation of optogenetic tools, calcium sensors, and other common neuroscience...for gut implantation. The fibers they ended up generating were 10-15 times more flexible than the brain...

...variant of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the...pAAV-hSyn-mCherry at Addgene. Viral vectors coming soon! These vectors listed below should be released at Addgene in the...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...the iGABASnFR and its F102G variant in AAV1 at Addgene! Archon1, a voltage sensor AAV Historically, electrical...Archon1 can also be used in conjunction with optogenetic activators and inhibitors that respond to blue... Find Archon1 and its cre-dependent version at Addgene! Fluorescent protein and control AAV These fluorescent...item is available for order please email help@addgene.org with the vector name and serotype you’d like ...

...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...Seeding Labs Resources on the Addgene Blog Listen to Additional Addgene Podcasts Learn How You Can Contribute...Contribute to the Addgene Blog Learn about Addgene Plasmid Donations for Rare Disease Research Resources...Resources on Addgene.org Become a part of the Addgene Community ...Biotechnology, he did his PhD in Plant Breeding, Genetics, and Biotechnology at Michigan State University.... Kwapata now works as a lecturer in molecular genetics and biotechnology for Horticulture and Forest ... recently received an award from nonprofit and Addgene collaborator, Seeding Labs, for lab equipment that...

...recently awarded plasmids from Addgene as a successful applicant to the Addgene, Seeding Labs Plasmid Grant...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...Resources on the Addgene Blog Check Out Posts on Synthetic Biology Learn about the Addgene, Seeding Labs...In this episode of the Addgene Podcast, we sit down with Kwabena Duedu, a researcher at the University... research experiences as well as plasmids from Addgene to bring new opportunities to Ghanaian scientists... biology needs and why Kwabena applied for the Addgene, Seeding Labs plasmid grant. 7:12 - 7:11 : Kwabena's...Sciences in Ghana. He is a recent recipient of the Addgene, Seeding Labs Plasmid grant and is working on a...

...Imaging Tools Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent... visualize their activity. In 2019, the first generation of norepinephrine sensors was developed, named...norepinephrine (Feng et al., 2019). But this first generation of tools had limited sensitivity and speed and... models of nLightG (left) and nLightR (right) generated using ColabFold (Mirdita et al., 2022). B, Representative...Speed One of the major advantages of this next generation of norepinephrine sensors is their speed. nLightG...GRABNE were more sensitive than nLightR, both generating a measurable signal after electrical stimulation...test point, ten pulses. This is in line with the general findings that red fluorescent proteins tend to ...

...found at help.addgene.org, and the widget is on any page on our regular site, www.addgene.org. We’re excited...comes first. Resources on the Addgene Blog Read other Inside Addgene Posts Learn about New Features ... at Addgene Visit the Help Center Find Plasmids for Your Research Become a Member of the Addgene Community...Addgene’s customer service philosophy Customer service can be a hassle. You finally make it through a... familiar? Well, when it comes to working with Addgene, we certainly hope not! One of our core values ...answer that you’re looking for, or you just have a general question or concern about the answers we provide...excited to make these new features available to the Addgene community, and we’d love to know what you think...

...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check out our CRISPR Featured...and designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found...cytosine deaminase. Three rounds of rational mutagenesis on the ADAR2DD residues that contact the RNA ...RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE...PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. “RNA targeting with...PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. "C2c2 is a single-... PMC5127784. Find plasmids from this paper at Addgene. East-Seletsky, Alexandra, et al. "Two distinct...

...share your lab protocols online Addgene Website: http://www.addgene.org/ If you’re working with plasmids...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...protocols. Since we began sharing this protocol on the Addgene website in December 2006, the pLKO.1-TRC cloning...the UK, South Korea, and beyond!   Check Out How Addgene Thinks About Developing Lab Management Software...plasmids and you decide to share them using Addgene’s repository, you can also share your associated protocols...submit an updated protocol? Just email us at help@addgene.org and we will be happy to upload your newest resources...gRNA_Cloning Vector (deposited by George Church’s lab). Addgene has also cataloged a variety of useful molecular...