...2007; plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have ... when using AAVs Resources on Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse all viral... neuronal cells. The AAV genome is a single-stranded DNA molecule and AAV vectors are replication deficient...step in the viral life cycle. Using AAV for neuronal tracing AAV vectors are the first choice of many neuroscientists... a cocktail of these different AAV vectors into a brain region, the AAV particles will infect neurons ...unique hue (plasmids at Addgene). A large-scale example of neuronal tracing with AAV vectors is the Allen... a new AAV variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV serotype...

...operator. AAV AAVs are the workhorses of both chemo- and optogenetics. We’re therefore beginning our AAV viral...have retrograde AAV that you can use to map neuronal connections. Expanding the viral vector knowledge base...Research: A Primer for Beginners Lentiviral Vector Uses and Overview AAV: A Versatile Tool for Gene Expression...spending time and money producing virus from select vectors in the repository yourself, you can now order ready-to-use...currently available) and adeno associated virus (AAV, primarily chemogenetics tools for now but with optogenetic...accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the years, many... production process while still ordering viral vectors. Virus production can be a frustrating process ...

...Viral Vectors 101: Viral Vector Elements Viral Vectors 101: An Introduction to AAV Viral Vectors 101:...Adeno-associated Virus (AAV) Guide Addgene’s viral plasmids and resources Addgene’s viral services: AAV Resources... Adeno-associated virus (AAV) is a small, single-stranded DNA parvovirus. AAV can infect humans and some...Recombinant AAV largely does not integrate, and the addition of a helper virus or plasmid allows it to... generally not as long lasting as AAV (up to several weeks vs. AAV’s several years). But these viruses...their required components are very similar. While plasmids containing virus-specific elements such as the...replicate and divide along with the endogenous DNA. AAV also has low immunogenicity, meaning it does not ...

...-stranded AAV (ssAAV) and a self-complementary AAV (scAAV) vector. Full-length ssAAV vector genome of ...about Viral Vector Elements Resources on Addgene.org Read Our AAV Guide Find AAV Vectors Browse All ...polarities are rescued from the vector plasmid and individually packaged into the AAV capsids. As a genome conversion...the basics of AAVs down, you’re ready to take the next step – selecting your AAV and AAV components (serotype...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...

...Looking for more viral vectors? Browse our in-stock viral vectors or consider our AAV Packaged on Request... 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...Addgene's Data Hub What is a Virus (and a Viral Vector)?  AAV Variables that Matter...

...micrograph of an Addgene AAV viral vector preparation after negative staining. Empty vector particles can be ...get more reliable AAV titering? For our viral service, we currently titrate AAV vector preparations by ...infectivity of wild-type AAV as benchmark for infectivity of recombinant AAV vectors." Gene therapy 17.7 (...Considerations When Using AAVs Addgene's AAV Quality Control Methods Check Out Our Viral Vector Featured Topic Page...to titer AAV! The point is there are many ways to quantify the composition of a viral vector solution,... viral vector preparations. Addgene has used this method to generally validate our viral vector preparation...darker than full vector particles. This image shows that the vast majority of the vectors consist of full...

...split vector strategies plus one additional split vector strategy for increasing the size of the AAV donor...homology directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the...time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an AAV transfer...transfer plasmid Resources on Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps ...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging capacity.... So how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces...into AAV First, some history. In 2008, the Auricchio lab decided to solve the issue of “tiny vector, big...

...scientists will be able to request an AAV composed of an AAV plasmid in the Addgene repository and their...titers for plasmids packaged in AAV1, AAV5, AAV8, and AAV Retrograde to be 1x1013 and titers for AAV2 to be...learn more.  What will be available?  Over 3000 AAV plasmids in the Addgene repository will be available ...Packaged on Request service. A small subset of AAV plasmids will have technical or legal restrictions that...recommend you check the original paper of your AAV plasmid of choice for more information on its performance...different serotypes for packaging: AAV1, AAV2, AAV5, AAV8, or AAV Retrograde (AAVrg) capsids.  Please be aware...excited to announce that we are expanding our viral vector service to include a Packaged on Request option...

... cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap...present in an AAV prep. AAV samples are first DNaseI digested to remove any residual AAV plasmid that was ...technique when handling your AAV plasmid standard and AAV transfer plasmids. It’s easy to contaminate a...My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, ...needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting HEK293...RepCap plasmid which provides the AAV replication (rep) and capsid (cap) genes. AAV replication uses the...that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus...

...Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral vectors ... better vector for ocular gene therapy: Adeno-Associated Virus (AAV) While other viral vectors such as...Adeno-Associated Virus (AAV) for cell and gene therapy AAVs for genome editing AAV: A versatile tool for...technician in a lab using adeno-associated viral vectors (AAVs) to treat an inherited retinal degenerative .... At the time, adenoviral vectors were the most commonly used viral vectors for gene delivery, and they...However new viral vectors capable of providing long-term gene expression in retinal cells - AAV - were becoming...are known to transduce retinal cells, AAV quickly became the vector of choice for ocular gene therapy. ...

...CRISPR/Cas9 and AAV Viral Vector Elements Additional Resources on Addgene.org Find AAV Vectors for Your Research...capacity, AAV vectors were suitable for our needs for the following reasons: Safety: Since AAVs are nonpathogenic... cells an AAV can infect) can influence an AAV’s value greatly. Applying this mantra to AAV selection,...viral-vectors/aav/aav-guide. Where in the tissue/what cell type? In the CNS, for instance, there are marked... I knew little about adeno-associated Viral Vectors (AAVs) before starting my graduate program at the ...types in locomotion. We have relied heavily upon AAV vectors to deliver chemogenetic/optogenetic constructs...nonpathogenic, we are able to use AAV vectors without specific facilities (Biosafety Level-1). Other possible...

...single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost... is a dual-vector system, meaning it is typically used in conjunction with a second vector that delivers...alternative single-vector systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast...constructs (i.e., using a dual-vector system as opposed to a single-vector system) has several benefits.... transductions with a single-vector system). Tips for using this plasmid Cas9 expression is typically ...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...

...together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...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...

...also called AAV2-retro) Looking for more viral vectors? Browse our in-stock viral vectors or consider ...purification, and titering. Our newest viral vector service, AAV Packaged on Request  has seen a fantastic...consider our AAV Packaged on Request service, which offers over 3,000 eligible transfer plasmids and five different...Addgene's Data Hub What is a Virus (and a Viral Vector)?  AAV Variables that Matter...serotype pENN.AAV.CMVs.Pl.Cre.rBG AAV5 Recombinase James M. Wilson New serotype AAVrg = AAV retrograde...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes pAAV-CAG-emiRFP670...Brain Science New viral tool pGP-AAV-CAG-FLEX-jGCaMP8m-WPRE AAV5 Biosensor GENIE Project New serotype...

...or on our hot plasmids webpage. Listen to the entire Hot Plasmids podcast! Hot plasmids covered in this...the Tol 2 Gateway-Compatible Toolbox  Hot Plasmid 2: AAV Vectors for tageting GABA-ergic interneurons. Listen... Listen to Our Segment on these new AAV Vectors Hot Plasmid 3: mCyRFP1 - a new, monomeric, and cyan-excitable... of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for targeting...nervous system, and vectors for zebrafish engineering. You can find additional hot plasmids in our newsletter...this segment Hot Plasmid 1: The Tol2 Gateway-Compatible Toolbox for quickly generating zebrafish transgenic... protein. Listen to Our Segment on mCyRFP1 Hot Plasmid 4: mScarlet - a new, bright, and monmeric red fluorescent...

...guide to viral vectors Find all of our viral vector blog posts Download our viral vectors 101 eBook Additional...ways to improve viral vector production. Whether redesigning viral packaging plasmids, investigating different...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes...improve their AAV yields. Given the wide range of downstream applications scientists rely on our AAV preparations...adapted by vector cores and laboratories alike as an easy and inexpensive method to improve AAV yields. ...pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering...To our knowledge the effect of hyperosmolarity on AAV production yields had yet to be examined. Increased...

...lab are now available as ready-to-use AAV vectors in serotype AAVrg.   ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...and putative pre-synaptic terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun...cortical interneurons. We now provide ready-to-use AAV vectors expressing GCaMP6f with this promoter. Image ...dendritic neurites from axonal neurites with AAV vectors expressing cytoplasmic targeted tdTomato and ...

...questions!  AAV Prep and Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential...compared to just one vector.   Q2: Do you think it is possible to make an efficient AAV prep for a 4.2 kB...the CMV promoter using the AAV-BR1 capsid? A: It is our understanding that AAV-BR1 has the same genome size...served by a dual vector, split intein approach.   Q3: What’s the best method for titering AAV? Is there a ...samples within a project. Targeting Your AAV Q4: Are there any AAVs that have successful transduction in ...different AAV particles into the same cells in vivo? A: This will be highly dependent on vector dose. The...Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page...

...choosing AAV or lentiviral vectors in our previous blog posts.  Viral vector injection Viral vectors allow...expression vector and the vector particles are delivered into the brain region of interest. AAV or lentivirus...lentivirus?  Both AAV and lentiviral vectors are used for intracerebral injections. AAV particles are smaller...lentiviral vectors are superior packaging vectors for larger genes - they can hold up to 8 kb, while AAV can ...viral vectors blog posts Find tips for preparing AAVs View our Beginner's Guide to Viral Vectors Resources...possible outcomes. Typically, AAVs have to be minimum around 1012-1013 vg/ml (vector genomes per milliliter)...using viral vectors By modulating neuronal gene expression and activity, you can use viral vectors to study...

...transfection of single plasmids vs pooled plasmids; for multiple edits, single plasmids are generally more...expression vectors which are often more difficult to transfect or package into viral vectors. Cas9 multiplexing... from crRNA expression plasmids resulting in smaller, more streamlined vectors which are easier to use...transduction, and AAV transduction.  In most of Zetsche et al’s experiments, a single vector co-expressing...transfected on a single plasmid. This is versus 2.4% of cells transfected with a pool of plasmids that contained...contained one crRNA per plasmid plus a Cpf1 expression vector.  See the graph in figure 2 for a comparison of...array that can be processed by Cpf1. Lastly, an AAV vector was used to multiplex edit 3-4 genes in a primary...