...needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting HEK293...present in an AAV prep. AAV samples are first DNaseI digested to remove any residual AAV plasmid that ... lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, I had... cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap...plasmid which provides the AAV replication (rep) and capsid (cap) genes. AAV replication uses the host’...the Addgene AAV Production protocol for more details. Pro tips I’m not going to lie, AAV transfer plasmids...experiment. AAV is usually the limiting reagent for my experiments, so I always make more AAV than I think...

...while both AAV and lentivirus have low immunogenicity, AAV still beats lentivirus. In mice, AAV is almost... vivo option AAV has become the preferred delivery method for in vivo CRISPR screens. AAV vectors can ...vivo CRISPR screens, AAV is unlikely to cause bad reactions in mice. Customization AAV offers a level of ...identified serotypes, AAV also has psuedotypes. Psuedotyping is the process of making a hybrid AAV, with the genome...degree using different types of AAV. Expression Another advantage of AAV vectors is how the cargo — in ...lentivirus, cargo delivered by AAV does not integrate into the host genome. AAV instead relies mainly on episomal...using AAV is the limited packaging capacity. Lentiviruses have capacities of 8–12 kb, while AAV only has...

...pure adeno-associated virus (AAV) preps for use in your research.... Protocols AAV Purification by Iodixanol...Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient Ultracentrifugation You may also...Introduction This protocol can be used to purify AAV of any serotype. In medicine, iodixanol can be used...performing a buffer exchange before using the purified AAV in vivo . The iodixanol gradient in this protocol...steps that separate out contaminants from an impure AAV preparation. The 15% iodixanol step has 1M NaCl to...containing) particles. On average, iodixanol-purified AAV preparations contain ~20% empty particles. 60% iodixanol...

...genome-containing particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s AAV Titration protocol. ... Protocols AAV Titration by qPCR Using SYBR Green Technology AAV Titration by qPCR Using SYBR Green Technology...validated using an internal reference AAV of known titer, 100837-AAV1, and by measuring the titer of samples...Virus Blog Posts Mol Bio Protocols Introduction The AAV Titration protocol can be used to determine the number... number of genome-containing particles of an AAV prep using SYBR green technology. This protocol was validated...samples in duplicate at least If possible, include an AAV reference sample of known titer. The reference material...when the plasmid is linearized. Treat the purified AAV samples with DNase I to eliminate any contaminating...

... launching the AAV Data Hub. The Data Hub is an open platform for scientists using AAV to share practical... rat. Contribute your AAV data Just like Addgene’s plasmid repository, the AAV Data Hub is a community-driven...Dear scientists using AAV, We hear your questions! How much virus should I inject? How long do I need...Hub and how to use it From the landing page of the AAV Data Hub, you will be able to search for your favorite...accompanying image so that you can see for yourself how the AAV performed in that experimental setup. For example...contribution to open science.  Please note that the AAV Data Hub is a beta project, and we are currently ...distributed by Addgene.  Future directions As the AAV Data Hub grows, we hope to mine the data to find ...

... Protocols ddPCR Titration of AAV Vectors ddPCR Titration of AAV Vectors You...You may also like... AAV Titration by qPCR AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular... additional tips on AAV titering using ddPCR, read this blog post on ddPCR for AAV quantification. This...for purified AAV titration. Protocols...purified recombinant Adeno-associated viral vectors (AAV). This protocol specifically uses primers and probes...series outlined in this protocol are based on an AAV titer range of 5E+12–5E+13 GC/mL, where GC is genome...pipettes and surfaces with 10% bleach. Safety Warnings AAV is generally considered biosafety level 1 but may...

...anti-AAV antibodies often neutralize the therapeutic efficacy of AAV vectors. How are exosome AAVs produced... exosome AAVs achieved so far? Exo-AAVs have improved transduction profiles in different AAV serotypes...lysis to release AAV particles with subsequent purification steps. It’s also possible for AAV vectors to be... They saw that these AAV particles come in a slightly different shape. These AAV particles are often found...name vexosomes (vector-exosomes), exosome AAVs, or exo-AAVs (Figure 1). The researchers discovered that... They also found that exo-AAVs were more resistant to neutralizing anti-AAV antibodies. This is particularly...microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol by Maguire...

...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid...Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory here...testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability...Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE 105322  AAV1 pGP-AAV-syn-FLEX-jGCaMP7c variant...Name 119741  AAV5  AAV SYN flex PSAM4 GlyR IRES EGFP Recombinase AAV Finally, we are excited to add...some of the AAV we have released in the last few months. You can also browse our entire AAV inventory. .... Our new AAVs include: Control AAVs for serotype testing jGCaMP7c for calcium detection PSAM4 GlyR for...

...types. AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV...-to-use AAV in Addgene's Viral Service Check out our AAV Production Protocol Check out our AAV Purification...wild type AAV is not associated with any pathogenicity and has a low immunogenic profile. AAV was first...of known AAV serotypes that have slight variations in their viral capsids. Novel forms of AAV capsids ...characteristic makes AAV vectors especially amenable to correcting disease-associated mutations. AAV vectors maintain... safe and effective. Challenges for AAV gene therapy Although AAV-based gene therapies are promising, ...already have wt AAV in their bodies and may therefore already have ways to prevent AAV infection. For ...

... cells an AAV can infect) can influence an AAV’s value greatly. Applying this mantra to AAV selection,...capacity, AAV vectors were suitable for our needs for the following reasons: Safety: Since AAVs are nonpathogenic...solution should be used to clean AAVs from workspaces. Speed: AAV transduction experiments are relatively...priority in our lab so we opted for AAV. While these benefits make AAVs useful for a wide variety of experiments...viral-vectors/aav/aav-guide. Where in the tissue/what cell type? In the CNS, for instance, there are marked...Serotype Striatum AAV5>AAV9>AAV8>AAV1>AAV6>AAV2 Hippocampus AAV9>AAV5>AAV8>AAV1>AAV6>AAV2 Auditory Cortex... use of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD ...

...can use AAVs to improve your genome editing experiments (with and without targeted nucleases). AAV genome...biology behind this AAV effect remains unclear, it is likely not a coincidence that the AAV genome is composed...disadvantages to the AAV-based approach that must be considered when deciding whether to use AAV or CRISPR for...of SaCas9 and a gRNA in one AAV and a repair template in second a second AAV to deliver their repair template...across two AAVs, it is possible sequentially repair a target site with the contents of the two AAVs. Using...to use and develop AAVs for your own research and encourage to deposit any new AAV tools you create. ...Resources on the Addgene Blog AAV: A Versatile Tool for Gene Expression in Mammals AAV Titers: Where do they ...

... serotypes AAV-hSyn-Ribo-jGCaMP8s AAV1 Biosensor Marianne Fyhn New viral prep AAV-hSyn-Ribo-jGCaMP8m...CN5140) AAV-PHPeB Optogenetics Jonathan Ting New viral prep pAAV_BiSSTe10_dTomato_nlsdTomato AAV-PHPeB...dTomato_nlsdTomato AAV-PHPeB Control Gordon Fishell New viral prep pAAV_BiVIPe4_ChR2_mCherry AAV-PHPeB Optogenetics...mCherry AAV-PHPeB Optogenetics Gordon Fishell New viral prep pAAV_BiPVe3_dTomato_nlsdTomato AAV-PHPeB ...pAAV_BiSSTe4_ChR2_mCherry AAV-PHPeB Optogenetics Gordon Fishell New viral prep AAVrg = AAV retrograde Looking...New viral prep pGP-AAV-syn-flex-iGABASnFR2(no bind)-WPRE AAV1, AAV5 Biosensor GENIE Project New viral...BGHpA (Alias: CN2237) AAV-PHPeB Control Jonathan Ting New viral prep AiP12787 - pAAV-AiE0140h_3xC2-minBG-SYFP2...

...makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids... want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let...experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV:  the packaging... the AAV transfer plasmid and take a look at each of its parts. Figure 1: Parts of an AAV transfer...   Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector. The cloning...ITR loss abolishing AAV packaging. To prevent ITR loss, it’s best to propagate AAV constructs in recombination-deficient...

...efficiency of AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for...efficiency of AAV-PHP.B versus AAV-PHP.eB in the CNS. A GFP transgene was packaged into either an AAV-PHP.B or...Search for AAV Vectors to use with your research. Visit Our AAV Guide Page for info on AAV components,... Recombinase-based AAV Targeted Evolution. In this technique, a pool of mutant AAV capsid (cap) expressing...astrocytes. Why screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but...one RGB AAV and red, green, or blue are the predominate colors expressed (figure 4c).   AAV Vector  ...et al.   AAV Vector Percent of Neurons Transduced   Dorsal Root Ganglion (DRG) AAV.9 (parental)...

...: pGP-AAV-syn-jGCaMP7f-WPRE (104488-AAV8) pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (104492-AAV8) pGP-AAV-syn-jGCaMP7c...)-EGFP (112677-AAV5) Calcium Sensors pGP-AAV-syn-jGCaMP7b-WPRE (104489-AAVrg) pGP-AAV-syn-FLEX-jGCaMP7b-WPRE...pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (104493-AAVrg) pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE (104495-AAVrg) pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE...some of the AAV we have released in the last few months. You can also browse our entire AAV inventory. .... Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors and GABA sensors Optogenetics...Optogenetics Recombinases And more! Control AAV For many AAV experiments, we recommend validating the serotype...samples of AAV for testing purposes. To address this, we recently released our serotype testing AAV, which...

...for a refresher on AAV titers Making contact with a cell Although an individual AAV particle’s ability...cell, the way to get more AAV in contact with the cell is to increase the AAV concentration. In this flowchart...optimize AAV production. We’ve listed some simple steps you can take below but you should read our AAV Production...on the Addgene Blog AAV Titers: Where do they come from and what do they mean? AAV Vector Quality Control... Find Ready-to-Use AAV Preps Find Lab Protocols for Work with Virus Watch Our AAV Purification Video ... place? Some AAVs are known to be difficult to produce in high titer. These include AAV2, which is known...Well, it turns out this isn’t the case for using AAV. When it comes to infecting cells, titer, the amount...

... parts of the AAV transfer plasmid  Resources on Addgene.org Learn about AAV Find AAVs for your experiment...making the AAVs you need for your experiment:  the packaging plasmid which contains the AAV structural...capsid (cap) gene. In wild-type AAV, these two genes are found between the AAV genome’s two inverted terminal...68 play a key role in AAV genome replication. Replication initiates from the AAV genome ITRs on the transfer...understanding of the AAV RepCap plasmid! Check out this blog post to learn more about the AAV transfer plasmid...terminal repeats (ITRs); but when recombinant AAV (rAAV) is made in the lab, rep and cap are expressed from...regulation of the AAV p40 promoter which drives Cap expression. Often Rep proteins from the AAV2 serotype are...

...by using the universal AAV Reference Standard Material (AAV SFM), which is an AAV sample that has been ... quantifying AAV: qPCR efficiency can be affected by second-order structure of the AAV genome, caused...we get more reliable AAV titering? For our viral service, we currently titrate AAV vector preparations ... et al., 2010). In addition to the AAV RSM, we include a second AAV reference sample of known titer, which... infer titer values of new AAV samples. We also typically quantify our AAV sample by two qPCR assays ...2017). What do Addgene AAV titers mean for your experiments? Because (physical) AAV titers are approximations.... If using AAV from different sources in the same experiment, consider retitering both AAV in your lab...

...the basics of AAVs down, you’re ready to take the next step – selecting your AAV and AAV components (serotype...single-stranded AAV (ssAAV) and a self-complementary AAV (scAAV) vector. Full-length ssAAV vector genome...chemogenetics experiments. Recombinant AAVs (rAAVs) The primary application of AAV is delivery of genetic cargo ...speak about AAVs, they are actually referring to rAAVs. ITRs and serotypes The names of AAVs contain two... lot of AAV serotypes, many of which are available in rAAVs!   Figure 1: Diagram of AAV components... self-complementary AAV (scAAV) may be an option (McCarty et al., 2001). This AAV is designed so that ...subtypes, since AAVs tend to be very neuro-permissive (Issa, 2023). Time to expression AAVs have to infect...

...injection site from the AAV data hub. How to share your AAV data Do you use Addgene AAV tools and want to... Using AAV Technologies Using AAV for neuronal tracing Resources on Addgene.org Read our AAV guide Browse...growing group of Addgene AAV data sharing champions? If you’ve recently used Addgene AAV tools, we invite you...contributed to the AAV Data Hub. If you haven’t yet, we encourage you to share your AAV data on the hub,... on AAV data hub! Related resources on the Addgene blog Important Considerations When Using AAVs Three... Browse all  AAV vectors Find ready-to-use AAV preps ... celebrate open data sharing than with Addgene’s AAV Data Hub! We launched this pilot in 2019 as an open...

...Gene Therapy. Existing methods for AAV quality control Historically, AAV vector QC has been sparse including...reliably confirm the identity of AAV genomes and detect contaminating AAV DNA. In addition, we wrote an ...the serotype of AAV preparations.  Using viral genome sequencing to determine serotype AAV packaging is ...LH (2017) AAV-ID: A Rapid and Robust Assay for Batch-to-Batch Consistency Evaluation of AAV Preparations...post on AAV titers Browse viral vectors protocols and tips Resources on Addgene.org Find AAVs at Addgene...platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed...VGS), to confirm the identity and serotype of our AAV preparations. We recently published a description...

... experiments with AAV: 1. Getting your viral vector into your cell of interest: AAV tropism and route ...administration AAV tropism is defined by the viral capsid proteins that determine the AAV serotype. During...characteristics of select AAV serotypes in Table 2 from the paper. Delivery of AAV into the brain can usually...our recently launched AAV Data Hub, an open platform where researchers using AAV from our viral service... Learn more about retrograde AAVs Resources on Addgene.org Read our AAV guide Learn more about our viral...With the help of adeno-associated viral vectors (AAV), neuroscientists can combine classical genetic tools...experiments! Late last year, we hosted our first AAV Education and Development workshop aimed at brainstorming...

.... Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! Optogenetics Optogenetics...pGP-AAV-syn-jGCaMP7b-WPRE 104493 AAVrg pGP-AAV-syn-FLEX-jGCaMP7b-WPRE 104496 AAVrg pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE ...Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg pGP-AAV-syn-jGCaMP7b-WPRE...some of the AAV we have released in the last few months. You can also browse our entire AAV inventory. ... control AAV in smaller aliquots of 20 ul for serotype testing to help you pick the best AAV for your ...26976 AAV5 pAAV-hSyn-hChR2(H134R)-mCherry 26973 AAVrg pAAV-hSyn-hChR2(H134R)-EYFP   Control AAV Control... 104495 AAVrg GP-AAV-CAG-FLEX-jGCaMP7s-WPRE 105714 AAV8 pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter ...

...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also ...Notes AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre...pAAV-EF1a-fDIO-Cre AAV5 Recombinases Esteban Engel New serotype pAAV-hSyn-Cre-P2A-dTomato AAV PHP.eB Recombinases...serotype pAAV-hSyn-hChR2(H134R)-EYFP AAV2 Optogenetics Karl Deisseroth New serotype AAV-CamKIIa-jGCaMP8s-WPRE...New viral prep pAAV-hSyn-GRAB-gDA3m AAV1 Biosensors Yulong Li New viral prep pGP-AAV-GFAP-iGABASnFR2...pGP-AAV-GFAP-iGABASnFR2-WPRE AAV1, AAV5 Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-GFAP-iGABASnFR2...

...questions!  AAV Prep and Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential... make an efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid? A: It ...our understanding that AAV-BR1 has the same genome size restrictions as other AAV. Therefore, one should...samples within a project. Targeting Your AAV Q4: Are there any AAVs that have successful transduction in ...animal models. All of our engineered AAV are presently built on an AAV9 backbone and so will very likely ...Did you catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If...however, that this will require co-delivery of two AAVs at a high MOI, often resulting in a drop in transduction...

...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...

...Addgene.org AAV Data Hub AAV Guide AAV Packaged On Request Browse all in-stock AAV preps Browse all AAV plasmids...β3 AAV8 Laminin receptor (LR)3 AAV receptor2   AAV9 Terminal N-linked galactose3 AAV receptor2... Figure 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1, AAV6, AAV7, and AAV8 in the...some AAV9 tools are available as in-stock preparations. AAV retrograde (AAVrg, also known as AAV2-retro...receptor Coreceptor AAV1 α2,3-/α2,6 N-linked sialic acids1 AAV receptor2   AAV2 Heparan sulfate ...

...    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...

...used viral vectors. That’s where AAV Packaged on Request comes in.  AAV Packaged on Request With Packaged...: The AAV Packaged on Request banner.   We currently offer serotypes AAV1, AAV2, AVV5, AAV8, or AAV...AAV Packaged on Request, an expansion of our viral vectors service, has officially launched! As of today...today, you can request any of over 3,000 eligible AAV cargo plasmids in the Addgene repository to be packaged...been following along, you may want to go to our new AAV Packaged on Request webpage to get started with your... go from selection to viral vector. To start the AAV packaged on request process, simply navigate to your...production and quality control.  Our packaged on request AAV service uses the same trusted production and quality...

...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...

... 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...

... 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...

...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...