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...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV_hSyn-SIO-stChrimsonR-EGFP-P2A-PdCO-miniWPRE...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the... the 20 additions we've made to our viral vectors repository since March.  Addgene is now offering an...This is an "optoGPCR" (coupled to Go, so it's an inhibitory opsin!) and due to its bistable nature, it enables... multiple serotypes Looking for more viral vectors? Browse the full viral services collection or check... check out Addgene's Data Hub to see viral vector reports from other users. And don't forget to use the...below to keep up with the latest in Addgene's viral vector services and resources!   This blog post was written...

...outside factors that need to be considered as well. In this post, we’ll touch on antibody storage, buffer....  Storage   Once you get your exciting new antibody, the second thing you’ll want to do is store it ...what temperature to store your antibodies at, but it also matters what they’re stored in - their buffer....the way, you can put your antibody in short term storage (think days to weeks) in the 4°C. If you’ll only...primarily used in Western blots and ELISAs, cannot be stored in sodium azide at all, as it inhibits HRP. In ...excitation by a laser, which is then read by a detector panel in the machine. Fluorophores are light sensitive...sensitive, so antibodies conjugated to them need to be stored either in a dark, opaque vial or wrapped in aluminum...

...blog  Viral Vectors 101: Biosensors Viral Vectors 101: Viral Applications Viral Vectors 101: An Introduction... an enzyme co-factor, a crosslinking agent, a signaling ion, and an allosteric modulator. Because of its...jGCaMP7 and jGCaMP8.   Viral vectors So, what do viral vectors have to do with all of these cool ...also already packaged into viral vectors and ready to go. Pro tip: Vectors with ready-to-use AAV preparations... a calcium sensor Genetically encoded calcium indicators (GECIs) are fluorescent proteins linked to calcium...troponin C). Their fluorescent readouts are used to monitor calcium concentrations and transients in living...living cells and organisms. Unlike chemical indicators, these DNA constructs can be delivered to intact tissues...

...blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Types of Viruses Viral Vectors 101: Viral...would not be great for a gene therapy vector! For viral vectors, only the protein for reverse transcriptase...components of the vector particle, some of the RT protein gets packaged inside of the vector particle, along...and this is where viral vectors and natural viruses diverge. For the viral vector, its job as a tool is ...Central PMCID: PMC372876 Dong, W., & Kantor, B. (2021). Lentiviral Vectors for Delivery of Gene-Editing Systems...seem like an unlikely candidate to use as a viral vector, but retroviruses have been developed into some...gamma-retroviruses — that have been developed into retroviral vectors and 2.) a subclass of retroviruses called the lentiviruses...

...Bicistronic vectors combine excitation and inhibition by using both excitatory and inhibitory channels activated...BioRender.com.   Opto-G-protein coupled receptors  G-protein coupled receptors (GPCR) regulate cellular function...some broad classes of optogenetic tools, and what factors to consider when choosing one.     Figure...light-gated ion channels, optogenetic G-protein coupled receptors (OptoXRs), and photoswitches. Light-gated ion ...stimulation to either activate or inhibit a specific receptor or pathway.  Light-gated ion channels Light activated...light within a specific range of wavelengths. Excitatory channels allow positively charged ions such as...gradient to depolarize it (Nagel et al., 2003). Inhibitory opsins are usually light-driven pumps that pass...

... on the Addgene blog Viral Vectors 101: Types of Viruses Viral Vector Elements Tips for Preventing... contact with than GFP. Similarly, if the viral vector carries an shRNA to a gene essential for cellular...cellular function, this would also be a higher risk vector. While it is unlikely that you, or anyone else ... you can receive appropriate treatment and/or monitoring. Virus-specific concerns   In nature, some viruses..., some adenovirus strains are known to cause respiratory diseases in humans, earning this virus a BSL-...

...Viral Vectors 101: Viral Vector Elements Viral Vectors 101: An Introduction to AAV Viral Vectors 101:...heterologous envelope or a post-transcriptional regulatory element can be used for either type of retrovirus...clinical settings. AAV is also a cool tool in the laboratory! It is ideal for whole animal administration,...population. Some adenoviruses are known to cause respiratory disease across species, as well as cancer in ...strains have been engineered to induce a lower inflammatory response within hosts. Relative to the other... Galla M, Baum C, Schambach A. Gammaretroviral vectors: biology, technology and application. Viruses. ...

...(scAAV) vector. Full-length ssAAV vector genome of both polarities are rescued from the vector plasmid...about Viral Vector Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All ...make AAV ideal for human therapies and in vivo laboratory experiments ranging from transient gene expression...a type 2 ITR and a type 8 serotype. Almost all vectors contain type 2 ITRs, which comes from the AAV2 ...capsid (outer protein shell) used to package an rAAV vector; these capsids allow the virus to infect (or not...multiple small genes, you can design single rAAV vectors with multiple genes packaged in them and use elements...or second strand synthesis. In contrast, a scAAV vector with half the size of the ssAAV genome has a mutation...

...Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors ...We offer a variety of viral vectors, viral services and even a Viral Vectors eBook to get you started. ... terrains. They are, in fact, a choice delivery vector from basic science research all the way to clinical... long-term, and even inducibly depending on the vector and virus. Viral CRISPR-Cas9 tools have been...interest, turning expression off permanently. Viral vectors can also be used to express proteins. Viral expression...of interest, and often a selectable marker. This vector can express your construct in target cells transiently...mechanisms require a specific modification of the viral vector for your gene of interest – an shRNA sequence specific...

...made to our viral vectors repository this year (and it's only March!)  In our repository, you'll find the...What's new in Addgene's ready-to-use viral vectors repository? Quite a bit! In this post, we'll share the...as seven vectors newly available in the AAV PHP.eB serotype. We've also added two viral vectors that can...these new vectors, and many more, in the table below.    Table 1: New ready-to-use viral vectors in the ...the Addgene repository  Name Serotype Category Depositor  Notes AAV-CAG-jGCaMP8s-WPRE AAV1 Biosensor...St-Pierre New viral vector pAAV-hSyn-JEDI-2P-WPRE AAV9 Biosensor St-Pierre New viral vector pAAV-CAG-RdLight1... Tian New viral vector pAAV-hSynapsin1-RdLight1 AAV9 Biosensor Tian New viral vector pAAV_hSynapsin_psychLight2...

...Patriarchi, T. (2023). Sensitive multicolor indicators for monitoring norepinephrine in vivo. Nature Methods...Neuromodulators like dopamine and norepinephrine have important functions in the brain but have been ...developing and depositing two new norepinephrine indicators: nLightG and nLightR (Kagiampaki et al., 2023...White insets indicate surface expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T...nLight sensors are based on alpha-1a adrenergic receptor; nLightG contains circularly permuted green fluorescent...trialed in two-photon in vivo experiments. nLight indicators are more sensitive to and selective for norepinephrine...This is important because alpha2-AR can provide inhibitory feedback in response to norepinephrine release...

... by sharing all the incredible guest posts and stories featured in our blog in 2023. Some of these were...Information DatabaseResearcher: Amos BairochViral Vectors 101: Systemic CapsidsResearchers: Cynthia Arokiaraj...

...recall, are the outer shell of an AAV vector, which determine the vector’s tropism, including its ability to...payloads. Since most systemic vectors are primarily derived from AAV9, the vectors have to be administered ...Hub and the CLOVER repository of published systemic AAVs to find commonly used vector cargo designs and... CNS (broad):  enhanced coverage for inhibitory versus excitatory neurons Mice, rats Yes (LY6A) ...AAV.CAP-B10 CNS: balanced coverage of inhibitory and excitatory neurons Mice, marmosets Unknown ...on the Addgene blog Viral Vectors 101: AAV Variables That Matter Viral Vectors 101: An Introduction to ...). Targeting AAV vectors to the central nervous system by engineering capsid–receptor interactions that...

...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV-CAG-emiRFP670...retro) Looking for more viral vectors? Browse our in-stock viral vectors or consider our AAV Packaged ...Addgene’s viral vector services allow scientists to conduct experiments faster, skipping the preparation...production, purification, and titering. Our newest viral vector service, AAV Packaged on Request  has seen a fantastic...serotype, and a dTomato under the control of the E6 regulatory element, which targets it to VIP cortical interneurons...Hub can help, with over 100 user-submitted viral vector reports. And don't forget to use the link below...below to keep up with the latest in Addgene's viral vector services and resources!   This blog post was written...

...the first base transversion editor The story of base transversion editors starts with researchers in the...Cytosine Base Editor Converts C → T; the first type of base editor invented. ABE Adenine Base Editor Converts...a glycosylase base editor (GBE). GBE Glycosylase Base Editor A base editor that relies on DNA glycosylation...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...transversion editors. Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows... depends on cellular factors. Multiple labs tried to identify beneficial factors to include in even better...DNA repair proteins to the base editor, they added the pioneer factor SOX2, which is capable of binding...

...researchers.  TALENs In 2010, transcription activator-like effector nucleases (TALENs) were engineered. TALENs... plague).  Base editors The major drawback of all the endonuclease-based gene editors (meganucleases, ...generate the prototypical base editor enzymes. The cytidine base editors (CBE) use a cytidine deaminase...the adenine base editors which can help accelerate the engineering of novel base editor proteins. She is... agricultural products. But it also made SciFi director Ridley Scott wonder what would happen if these...year after the discovery of meganucleases. The storyline involves the resurrection of dinosaurs by extracting...sequences is not ideal. For the early generation gene editors, new amino acid recognition sites had to be designed...

...reversion of this mutation by an adenine base editor would restore antibiotic resistance. To find such a protein...not require any additional inhibitor protein like UGI in CBE. Base editors for the plant genome These ...2: Plant base editor platforms and their molecular components. Adenine base editors for plant We developed... developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...smaller vectors suitable for transient assays using protoplast transformation. Cytosine base editors for ...plant base editors. Plant CBE and ABE with expanded PAM compatibility All of the base editors mentioned...require other base editor platforms. Hua et al. constructed a number of plant base editors replacing the SpCas9...

...plasmids use Rho-independent terminators, which include naturally occurring terminators, such as T7 and rrnB,...engineered terminators come close (>95%). For most purposes, however, any common terminator will suffice...suffice. Many commercial expression vectors use double terminators to reduce unwanted translation of downstream...require binding of termination factors. Although both are termination factor dependent, Polymerases I and...Posttranscriptional Regulatory Element Enhances Expression of Transgenes  Delivered by Retroviral Vectors. Journal...of yeast terminators of varying strength. A discussion of the effects of various terminators in the context...termination and polyadenylation? The role of the terminator, a sequence-based element, is to define the end...

...block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial constitutive...research purposes, retroviral vectors have been developed into ‘split-vector systems’ by separating viral...and expression of these vectors when used in experiments. Retroviral vectors make up a large toolbox ...Gamma-retroviral vs. lentiviral vector systems Gamma-retroviral vectors have a relatively high transduction... many more useful viral vectors available at Addgene: Follow our viral vector blog posts for updates on...tips for using retroviral vectors in your experiments. Gamma-retroviral vectors Popular gamma-Retroviral...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...

... download the PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging ...corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier to plan...for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting...right, you need to do a few calculations. This calculator can do the math for you and it’s simple to make...whole experiment And here’s the equation the calculator uses to give you the μl needed of each plasmid...experiments. Let us know what you think about the calculator in the comments section below. References 1....Production of Recombinant Adeno‐Associated Viral Vectors for In Vitro and In Vivo Use." Current Protocols...