... Adeno-associated virus (AAV) is a small, single-stranded DNA parvovirus. AAV can infect humans and some... generally not as long lasting as AAV (up to several weeks vs. AAV’s several years). But these viruses...Adeno-associated Virus (AAV) Guide Addgene’s viral plasmids and resources Addgene’s viral services: AAV Resources...currently known to cause any diseases. Recombinant AAV largely does not integrate, and the addition of a...replicate and divide along with the endogenous DNA. AAV also has low immunogenicity, meaning it does not ...conducive to targeting all cell types. Altogether, AAV has many excellent features for human gene therapy...and is currently being used in clinical settings. AAV is also a cool tool in the laboratory! It is ideal...

...multiple AAV variables.    Serotypes Serotype refers to the different protein compositions of AAV capsids...when designing and testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9...Alternate Route for Adeno-associated Virus (AAV) Entry Independent of AAV Receptor. Journal of Virology, 92(7)...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: The AAV Retrograde Serotype Viral Vectors 101...Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst... in and start injecting your virus, a successful AAV experiment starts with validation and optimization...usually faster and easier to do the validation first.  AAV-mediated gene expression can differ extensively depending...

... and quality control are described below. AAV Production AAV distributed by Addgene has been produced ...purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV subjected...arrowhead). Scale bar = 100 nm.   AAV resources Browse AAV preps available from Addgene. After receiving...Addgene blog Find and share AAV Data with Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors...Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at ...84445-AAVrg).   Electron Microscopy The ratio of empty to full (i.e., genome-containing) AAV particles..., we’ll walk you through how we produce both our AAV preps and our lentiviral preps.   Viruses are generated...

...Alternate Route for Adeno-associated Virus (AAV) Entry Independent of AAV Receptor. J Virol 92. https://doi.org...Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting Using AAV Technologies Resources... on Addgene.org Find ready to use AAV vectors at Addgene. Find AAV plasmids to use in your experiments...of viruses used (100798-AAV, lot #v15287, titer 1.7x10e13 GC/mL and 100799-AAV1, lot # v14715, titer 2.4x10e13...their cells of interest and recombinase-dependent AAV vectors for selective transgene expression. While... or spurious expression of recombinase-dependent AAV vectors. In this article, we will discuss some of...Fischer, KB. 2019). This means that all DIO/FLEx AAV vectors will have some level of recombination that...

...operator. AAV AAVs are the workhorses of both chemo- and optogenetics. We’re therefore beginning our AAV viral...currently available) and adeno associated virus (AAV, primarily chemogenetics tools for now but with optogenetic...researcher Connie Cepko has been helping us with AAV testing and recently had this to say about the service...preparations to the scientific community. Addgene's AAV preparations have worked well in my lab, including...ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can always head... will be available soon. We’re also distributing AAV preps with particular serotypes that can be used ...You can use these as controls to determine if your AAVs are getting to the right cells using your particular...

...YouTube page! AAV purification by iodixanol gradient ultracentrifugation This video goes over AAV vector purification...  Check out the AAV Purification protocol page for more detailed instructions. AAV titration by qPCR...service in 2016, Addgene has shipped thousands of AAV and lentiviral preps to scientists around the world...strategy to confirm the identity and purity of all AAV viral lots. Aligning with Addgene’s mission to “...your qPCR to determine the genome copies in a given AAV preparation. For detailed instructions and calculations...stock used to generate a standard curve, visit the AAV Titration by qPCR written protocol page. Getting...

...also offered as AAV1 (along with AAV PHP.eB, AAV Retrograde, AAV2, AAV5, AAV8, and AAV9). Once activated... the Dolcetto lentiviral prep here Control AAV Control AAV allow researchers to not only interpret their...offer control AAV in smaller aliquots of 20ul for serotype testing to help you pick the best AAV for your ...Testing AAV inventory for the entire collection! Plasmid Serotype Name Depositor 27056 AAVrg pAAV-Ef1a-DIO...ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released in the last...confirm expression in target cells. While the various AAV serotypes can be used to achieve different patterns...specific manipulation of neurons. We have expanded our AAV collection to include two new ChR2 constructs that...

...Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols...Overview of key steps of AAV production, iodixanol gradient purification, and AAV titration by qPCR.   Pro... addgene.org  AAV Titration by qPCR Using SYBR Green TechnologyAAV Titration by ddPCRAAV Purification ...Addgene AAV Production protocol. Titering Once you’ve produced, harvested, and purified your rAAV, you’ll...transducible units in the virus preparation.  rAAVs  Now your AAV virions are ready to store or use. They are...process.  Helper-free production Unlike lentivirus, AAV does not possess the ability to replicate itself ... applications, you can consider using unpurified AAV from the supernatant (Goodwin et al., 2020) or crude...

...adeno-associated viruses (AAVs), will know just how tricky these tools can be to use. AAVs, which can target ... Cumulative AAV preps available in the Addgene repository each year from 2016–2023. B) AAV preps distributed...: A) Cumulative AAV preps available at Addgene from 2016–2023 by type of tool. B) AAV preps distributed...before selecting an AAV can help a researcher make a more informed choice on which AAV to request and/or...feedback on AAVs at Addgene? Suggest a viral vector for our service; leave a comment; or take our AAV survey...the retrograde AAVrg and the systemic capsid PHP.eB.   Figure 4: A) Cumulative AAV preps for the...from the lengthy process of validating and using an AAV in a specific experimental setup. Teams across Addgene...

...production. AAV and lentiviruses can be produced in insect cells, but they require cloning specific AAV or lentiviral...cells Insect cells Viruses AAV and lentivirus Baculovirus; can produce AAV and lentivirus if needed ...Ultracentrifugation (AAV); centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti...Addgene blog Viral Vectors 101: Producing Your rAAV AAV Titers: Where Do They Come From and What Do They... and widely available. The viruses Lentiviruses, AAV, and adenoviruses can all be produced in mammalian... it is much more common to make lentiviruses and AAV in-house. High-titer adenoviruses are difficult and...production labs or companies. Production of both AAV and lentivirus requires three plasmids — a transfer...

... Protocols AAV Production in HEK293T Cells AAV Production in HEK293T...adeno-associated virus (AAV) in HEK293 cells. Protocols...HEK293T Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology...Introduction This protocol can be used to produce AAV from one Five Chambers Cell-Stack (CS5) (Link opens... Vortex Reagents Adherent HEK293T cells (ideally AAV-293T clones) Pro-Tip While adherent, these cells ...not use filters made of materials other than PES. AAV particles stick to many other surfaces, but do not...health of the HEK293T cells is critical for optimal AAV yield. Do not overgrow your cells. Pass the cells...

...powerful tool for quantifying AAV for some time now. When titrating AAV by qPCR, viral DNA is amplified...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ... is where droplet digital PCR (ddPCR) comes in. AAV titering using quantitative PCR Before we dive into...reactions with known quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used...and both results would be valid. This is why the AAV reference is critical. It allows you to determine...reference, meaning saving on reagents (and time)! AAV titering using droplet digital PCR Droplet digital... for your PCR experiment. The process to titrate AAV by ddPCR begins with diluting the virus. It is important...

...Addgene’s first AAV Data Hub Challenge, now taking place Dec. 6, 2022! Share your AAV data, connect with...with other researchers working with AAV, support open science, and maybe even win a $500 travel prize ...can submit data from their CLOVER lab or Addgene AAV experiments to the Data Hub. Submit your data by ... competition, where you can get all your burning AAV questions answered.  Interested? You can learn more...

...in the lab Over-Agar Antibiotic Plating Protocol AAV Purification Purify adeno-associated virus from a... a preparation produced in mammalian cells AAV Purification by Iodixanol Gradient Ultracentrifugation ...workspace, and maintaining sterility while working. AAV Titration by qPCR Use qPCR to measure the physical...titer of your adeno-associated virus preparation. AAV Titration by qPCR Using SYBR Green Technology Protocol...

...improve their AAV yields. Given the wide range of downstream applications scientists rely on our AAV preparations...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes...To our knowledge the effect of hyperosmolarity on AAV production yields had yet to be examined. Increased...sorbitol. As sorbitol is already a common excipient in AAV formulation buffers, this sugar may be a more attractive...fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference between...alike as an easy and inexpensive method to improve AAV yields. Although we only investigated 3 serotypes... approach will be beneficial for a host of other AAV serotypes; it will be exciting to test this approach...

...self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand AAV (ssAAV). To ...Voltron2 voltage sensors AAV vectors for targeting microglia The REVeRT Dual AAV Vector System Neurodegeneration... incorporated in an AAV vector (Figure 5). Efficiency and specificity of these AAV vectors for microglia...Dual AAV Vector System By Vaibhav Kawde To circumvent the payload limitations of traditional AAVs, the...the investigators found that administering these AAVs with a dilute Cre-expressing vector resulted in ... and highly specific targeting of microglia with AAV By Alyssa Neuhaus Genetic modification of microglial...CC-BY-NC-ND license.  Integrating hIBA1a promoter into AAV vectors, either alone or in combination with miR124T...

...about Addgene's 2018 AAV Education and Development (AAVED) workshop series....Development (AAVED) Series AAV Technologies and Methods for Targeted Neuronal Manipulation Meeting Date September... The AAV Education and Development...September 21, 2018 Contact Us Questions? Contact us at AAVED@addgene.org Sponsor We are grateful to The Kavli...brainstorm practical strategies for using combinations of AAV-based molecular tools to achieve targeted neuronal...Fan 9:30 AM Cre and Flpo Katherine Matho 10:15 AM AAV Retrograde Tracing Gowan Tervo and Sarada Viswanathan...Promoter/Enhancer Specificity, Serotype specificity, AAV dosing, and Other Forms of Regulation Ben Deverman...

...single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost...of the human synapsin promoter. Purpose of this AAV pAAV-hSyn-DIO-hM4D(Gi)-mCherry can be used to induce...ready-to-use lentivirus and adeno-associated virus (AAV) to scientists around the world. We began with only...expanded our viral inventory to 25 lentiviruses and 25 AAVs. These viruses have been distributed in over 200...again) to find the most requested lentivirus and AAV of 2016. The top viruses of 2016 were (drumroll please...transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain....Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our Viral Service Resources on ...

...week, we launched our new AAV Data Hub, an open platform for scientists using AAV to find and contribute ... posts Learn more about AAVs Resources on the Addgene website Visit the AAV Data Hub Learn more about...about the viral service Read our AAV guide ...last year on sucrose supplementation to increase AAV2 yields and we’re eager to share more! Lastly, we...

...concentrations or AAV titer for the best results. The timing of sacrifice when using Tetbow AAVs - You will ... #104105) using in utero electroporation or with AAVs (prepared with Addgene #104110 - #104112) (Figure...sacrifice the animals and fix the samples. When you use AAVs, it typically takes a few weeks to achieve optimal...your Tetbow experiment Plasmid concentrations and AAV titer - The XFP genes have to be introduced at ~2...achieve an adequate expression level of the Tetbow AAVs. However, you should not wait too long, as too much...Addgene blog Read about Brainbow Learn about using AAVs for neuronal tracing Visit the viral vectors blog...service See our fluorescent protein collection Find AAVs at Addgene ...