... Protocols AAV Production in HEK293T Cells AAV Production in HEK293T...HEK293T cells from 2 x T-175 flasks. Cells should be at ~80% confluence. For each T-175 flask: Aspirate ...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...

...H., Morin, E., Nguyen, T. N., Garren, E., Bendrick, J. L., Kim, T. K., Zhou, T., Mortrud, M., Yao, S.,...    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 ...

...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 ...

... 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...calculate the titer for each sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume...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...

...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..., J. T., Hantman, A. W., Looger, L. L., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV Variant...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...

...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... Byrne, B. J., Caton, T., Chiorini, J. A., Chtarto, A., Clark, K. R., Conlon, T., Darmon, C., Doria, M... 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 ...

...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... ITR sequences from different AAV serotypes that vary in their RBE and T-shaped hairpin loop sequences...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...

...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...

...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...

.... Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! Optogenetics Optogenetics...124603 AAV9 pAAV-EF1a-DIO-ChrimsonR-mRuby2-KV2.1-WPRE-SV40 124650 AAV9 pAAV-CamKIIa-C1V1(t/t)-mScarlet-KV2.1...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...

...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...

...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...travel award. All participants will receive an Addgene t-shirt.  Our judges, Drs. Connie Cepko, Ian Wickersham...

...37825-AAV5.T 20 µL $ 140 Add to Cart AAV8 37825-AAV8.T 20 µL $ 140 Add to Cart AAV9 37825-AAV9.T 20 µL $ 140...50465-AAV5.T 20 µL $ 140 Add to Cart AAV8 50465-AAV8.T 20 µL $ 140 Add to Cart AAV9 50465-AAV9.T 20 µL $ 140...Price* AAV1 37825-AAV1.T 20 µL $ 140 Add to Cart AAV2 37825-AAV2.T 20 µL $ 140 Add to Cart AAV5 37825-...Price* AAV1 50465-AAV1.T 20 µL $ 140 Add to Cart AAV2 50465-AAV2.T 20 µL $ 140 Add to Cart AAV5 50465-... Viral Vector Packaging Service AAV Serotype Testing AAV Viral Vector... Vector Packaging Service: Serotype Testing AAV These AAV encode fluorescent reporters and can be used...experiments. For more information, see our full AAV inventory or our AAV production page . Note: For a list of ...

...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...

...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...., Isa, K., Mizukami, H., Ozawa, K., Isa, T., & Yamamori, T. (2015). Comparative analyses of adeno-associated...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...

...transcription. Thus, T-DDOGs can be used in combination with green-red Cre-lox systems. T-DDOGs can also be...not red fluorescent proteins, as seen with T-DDOGs. Like T-DDOGs, Cre-DOG is both robust and specific...and 100% of DsRed+ cells also expressed GFP. Using AAV constructs, Tang et al. found that Cre-DOG could ...optogenetic studies, and since cells infected with AAV-Cre-DOG retain normal neuronal function, this system...only in the presence of GFP. These systems, known as T-DDOG and Cre-DOG, respectively, repurpose popular ...luciferase transcription. Tang et al. coined this system T-DDOG (Transcriptional Devices Dependent On GFP). ...et al. found that YFP and CFP can activate certain T-DDOG constructs similarly to GFP, but commonly used...

...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...

...C1V1 (t/t)-TS-EYFP EF1a C1V1 (t/t) EYFP Cre dependent 9 Deisseroth 35499 pAAV-CaMKIIa-C1V1 (t/t)-TS-EYFP...TS-EYFP CaMKII C1V1 (t/t) EYFP Constitutive 9 Deisseroth 124650 pAAV-CamKIIa-C1V1(t/t)-mScarlet-KV2.1 CaMKII...pAAV-Ef1a-DIO C1V1 (t/t)-TS-mCherry EF1a C1V1 (t/t) mCherry Cre dependent 9 Deisseroth 105448 pAAV-hSyn-DIO-ChrimsonR-mRuby2...Browse all Optogenetics AAV *AAVrg = retrograde serotype, produced with the AAV retro helper plasmid from...high quality AAV preps from select plasmids in the repository. Browse our optogenetics AAV collection, ...Packaging Service AAV Optogenetics Viral Vector Packaging Service: Optogenetics AAV Optogenetic tools ...CaMKII C1V1 (t/t) mScarlet Constitutive 9 Harvey 59170 pAAV-Syn-Chronos-GFP Syn Chronos GFP Constitutive...

...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...

...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 ...post on AAV titers Browse viral vectors protocols and tips Resources on Addgene.org Find AAVs at Addgene...Barungi T, Penaud-Budloo M, Broucque F, Blouin V, Moullier P, Ayuso E, Vandenberghe LH (2017) AAV-ID: A ...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...