...together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this...co-infection of two AAVs. Cas9 orthologs: shorter, but just as potent and specific? The two AAV strategies described...for in vivo studies. Testing AAV-SaCas9 in vivo To test the efficiency of AAV-SaCas9 in vivo, the Zhang ...not increased compared to a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs mediate genome...proteins with AAV, at just 1,108 and 986 amino acids, respectively, the size of both is within AAV’s packaging...in length, allowing them to be packaged in AAV.   Split AAV approaches  While finding smaller Cas9 orthologs...that the field has overcome with split AAVs. In general split AAVs break a large transgene in two pieces...

...and Share AAV Data with Addgene's New AAV Data HubCelebrate Open Data Day with Addgene’s AAV Data Hub!...with AAV vectors in neuroscience Let’s consider my first challenge when I started working with AAV vectors... answers hundreds of AAV questions every year on a range of issues related to AAV vectors, many from people...was: What AAV vector is best for my specific system?  Sharing data and community concerns AAV vectors ...transfer to the rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(... Fig. 1: Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2 noradrenergic...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists...

...useful, DREADD-containing AAV here. But the top requested AAV that became available in 2017 is pmSyn1-EBFP-Cre...specifically in neurons. AAV Delivered Cre - A useful tool for neuroscientists AAVs are widely used by neuroscientists...Hongkui Zeng’s lab for providing this AAV vector! Additional Resources AAV Titers: Where do they come from...service is going strong, the AAV service has shown incredible growth this year. pAAV-hSyn-DIO-hM4D(Gi)-mCherry...This AAV has had over 150 orders since coming online! pmSyn1-EBFP-Cre Available in serotypes AAV1, AAV5...mCherry was the top requested AAV prep for the 2nd year running, and you can learn more about this useful...AAV5, and AAV retrograde (the most popular form), this viral vector is used to deliver DNA encoding Cre...

... serotype pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE AAV PHP.eB Biosensor Kim New serotype AAV-mDlx-jGCaMP8s-WPRE...Notes AAV-CAG-jGCaMP8s-WPRE AAV1 Biosensor Looger New serotype AAV-CamKIIa-jGCaMP8m-WPRE AAV5 Biosensor...vector pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls Wilson New serotype pAAV-CaMKIIa-EGFP AAV PHP.eB...pAAV-mDlx-NLS-mRuby2  AAV PHP.eB Controls Gradinaru New serotype pAAV-CaMKIIa-mCherry  AAV PHP.eB Controls Deisseroth... AAV-hSyn-Soma-jGCaMP8f AAV9 Biosensor Fyhn New serotype pAAV-hSyn-JEDI-2P-Kv-WPRE AAV9 Biosensor St-Pierre...-WPRE  AAV9 Biosensor Looger New serotype AAV-hSyn-Soma-jGCaMP8s AAV9 Biosensor Fyhn New serotype ...Chemogenetics Deisseroth New viral vector pAAV.CMV.PI.EGFP.WPRE.bGH  AAV PHP.eB Controls Wilson New viral vector...

...Using AAVs AAVs Are Great Tools for Gene Expression in Mammals Resources on Addgene.org Find AAV Packaging...PEI transfections for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires... to making AAV packaging a little easier! Click here to download the PEI calculator for AAV packaging ...calculator for AAV packaging Select the plasmid molar ratio you would like to use for your AAV packaging ...Making AAV? Check out Addgene’s Step-By-Step Protocol Pressed For Time? Order Ready-to-Use AAV from Addgene...this calculator will make it easier to plan your AAV packaging experiments. Let us know what you think... aav...

...    Figure 1: AAV toolkit for targeted gene delivery to the brain. The AAV (a) capsid, (b) cargo...precious cargo space in AAV vectors. Dual expression systems, which use two or more AAVs to transduce the same... et al, 2019 AAV-PHP.B2 CNS Mice Yes (LY6A) Deverman et al., 2016 AAV-PHP.B3 CNS Mice...2022 AAV-PHP.eC CNS: neurons and astrocytes Mice No (LY6C1) Shay et al., 2023 AAV-PHP.V1...al., 2020 AAV-PHP.N CNS: neurons Mice Yes (LY6A) Kumar et al., 2020 AAV-PHP.B4 CNS ...2023 AAV-F CNS: neurons and astrocytes Mice No (LY6C1)   Hanlon et al., 2019 AAV-BI-28 ...from other labs. AAVs not otherwise attributed were deposited by the Gradinaru lab.  AAV-PHP.B was the first...

...time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an AAV transfer... of ~9 kb into AAV. They found that in defiance of the packaging limit dogma, this AAV successfully mediated...size of the AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches...independently packaged into an AAV. When a cell is co-infected with both part A and part B AAVs, the fragments reassemble...Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral service ...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging ...how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces. Reassembly...

...Adeno-Associated Virus (AAV) for cell and gene therapy AAVs for genome editing AAV: A versatile tool for...pigmented epithelium (RPE). AAVs, on the other hand, have broad tropism in the eye. AAV-2, the first identified...occurring and engineered AAV serotypes that further expands the range of AAV properties is available. ...mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral vectors...into the field of gene therapy, more specifically AAV-based retinal gene therapy. The year was 2001 and...technician in a lab using adeno-associated viral vectors (AAVs) to treat an inherited retinal degenerative disease...and cell) therapies. One may wonder why the first AAV-based gene therapy treatment for a genetic disease...

...Our new AAVs include: pGP-AAV-syn-FLEX-jGCaMP7b-WPRE pAAV.hSyn-FLEX.iGABASnFR pAAV.hSyn-FLEX.iGABASnFR.... of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the primary...are some of the AAV we have released so far in 2019. You can also browse our entire AAV inventory! Our ...Plasmid Serotype Name 50363  AAV5  AAV phSyn1(S)-DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre (FLP-dependent...Plasmid Serotype Name 104496  AAV1  GP-AAV-CAG-FLEX-jGCaMP7f-WPRE AAV1 with a CAG promoter exhibits high...iGABASnFR and its F102G variant in AAV1 at Addgene! Archon1, a voltage sensor AAV Historically, electrical activity...114472  AAV5, AAV8  pAAV-hSyn-mCherry 50459  AAVrg  pAAV-hSyn-DIO-mCherry 59462  AAV2  pAAV-CAG-tdTomato...

...Performance of synthetic promoter AAVs in mice All of these synthetic promoter AAVs were first injected into the...  Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular cell... A toolkit of synthetic AAV promoters The Roska lab’s synthetic promoter AAVs allows for targeted gene...about using AAV for retinal gene therapy Find more about precise neuronal targeting using AAV technologies...posts Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral vectors...Adeno-associated virus (AAV) is a popular tool for gene delivery, but it has a drawback: how do you ensure...basic research. To improve cell-type specificity of AAV, work has focused on modifying the outside protein...

... How did AAV-68544 become contaminated? Well, we purposely mixed another AAV vector with AAV-68544 to ...identified, and we will discard any AAV prep we suspect is contaminated. For AAV-68544, the best match for the...). When we blindly analyzed AAV-68544, we were easily able to identify AAV-44632, the contaminant we mixed...because of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake ...research. To make sure that adeno-associated viral (AAV) vectors available at Addgene are suitable for in...perform all the standard quality control assays for AAVs. These steps include measuring viral titer and assessing...Why should you care about viral quality control? AAV vectors are produced by scientists - not robots. ...

...expression.     AAV retrograde Unlike traditional dye-based neuronal tracing reagents, AAV particles require....)  In this post we’ll describe the AAV retrograde serotype (AAVrg) and how it enables scientists to access...look how cool the brain is!" Using retrograde AAV AAVrg can be used to deliver a wide array of genetic...inhibited) by any genetic tool delivered by the AAV.   AAVrg requires high titers for efficient retrograde... can find in AAV Data Hub!) The cortical pathways to the IC were well transduced by AAVrg while other ...difficult. The in vivo workhorse adeno associated virus (AAV) shows relatively low retrograde transport efficiency...University of California, Berkeley engineered an AAV variant that efficiently infects neurons, directs...

...and re-launched Addgene AAV Data Hub, the place for scientists to share their AAV validation data. The Data...scientists, learn more about the AAV Data Hub, and hear the incredible AAV talks given by Connie Cepko and... of community submissions for Addgene and CLOVER AAV experiments - and we've updated our submission process...Using the Data Hub If you're trying to decide which AAV is right for your experiment, searching the Data ...can even see dendritic spines!  Curious? Watch our AAV Data Hub Challenge recording to see two reports presented...Hub If you've ever validated an Addgene or CLOVER AAV, you can (and dare we say, should!) submit your data...meet FAIR data-sharing guidelines. But what if my AAVs didn't work?  We love negative data, and in fact...

...nervous system (PNS) targeting AAV  Get 1st time AAV user tips from an AAV rookie  Learn about some considerations...Search for AAV vectors to use with your research. Visit Our AAV Guide Page for info on AAV components,...transfer to the rat CNS: AAV PHP. EB vector dose–response and comparison to AAV PHP. B." Gene therapy 25.5...considerations when using AAVs Learn more about using AAV as a tool for gene expression in mammals. Resources...components, common uses of AAV, and much more! Find ready-to-use AAV from Addgene or check out protocols for packaging... its parental AAV9 relied on for transducing cells: galactose and the AAV receptor (AAVR). Cell lines ...such as adeno-associated virus (AAV), from reaching their targets. The AAV9 serotype can breach the BBB ...

...considerations when using AAVs Resources on Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse ... a new AAV variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV serotype...plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have been widely...into neuronal cells. The AAV genome is a single-stranded DNA molecule and AAV vectors are replication ...proteins. For a brief overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size... CNS AAV1, AAV2, AAV4, AAV5, AAV8, AAV9 Heart AAV1, AAV8, AAV9 Liver AAV7, AAV8, AAV9 Lung AAV4... step in the viral life cycle. Using AAV for neuronal tracing AAV vectors are the first choice of many...

...are the elements that make an AAV plasmid an AAV plasmid.  Within the AAV genome, ITRs serve as the origin...capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids from multiple AAV serotypes...the single stranded DNA genome of AAV in the native virus. In an AAV vector plasmid, they flank the gene...ITRs are retained with the gene in a recombinant AAV (rAAV) vector (Figure 1). Everything else outside the...ITRs are one of the most important elements of AAV and rAAV vectors, modifications can drastically affect...Adeno-associated viruses (AAV) are an important biological tool that allow scientists to express a gene...gene of interest in a cell or animal model. AAV plasmids contain several important elements, but among...

...release from late endosomes when AAV is trafficked. Rep and cap mediate AAV pseudotyping which allows for...for altered tropism of AAV to target different tissue or cell types. AAV also requires some back up from...vector components of AAV Adenovirus Adenoviruses have quite a bit in common with AAV, just like lenti ...there are many shared components across viruses (AAV, lentivirus, etc.) and their functions are more intuitive...lentivirus, adenovirus, and adeno-associated virus (AAV). The components of these vectors are also split ...components of retrovirus  Adeno-associated virus AAV’s elements are quite different from gamma-retrovirus...cell and deposit viral cargo. Gene expression While AAV often does not integrate into the host genome (more...

... 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...