...lentiviral plasmids Addgene's most popular retroviral plasmids Addgene's collection of AAV plasmids Addgene...adeno-associated virus (AAV). The components of these vectors are also split across several plasmids (often three... Viral vector components of AAV Adenovirus Adenoviruses have quite a bit in common with AAV, just ...adenovirus Building viral vectors Addgene has many viral vectors and helper plasmids for all the viruses discussed...of adenovirus plasmids Resources on the Addgene blog Parts of the AAV packaging plasmid Overview of Lentiviral...many plasmid maps and protocols! In reality, there are many shared components across viruses (AAV, lentivirus...of viral vectors here! Need Virus? Check out Addgene's Viral Service! Types of viral vectors Of the many...

...control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid Serotype...sensors Plasmid Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE 105322  AAV1 pGP-AAV-syn-FLEX-jGCaMP7c...can also browse our entire AAV inventory. Our new AAVs include: Control AAVs for serotype testing jGCaMP7c...Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory here... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...recombinases Plasmid Serotype Name 55637  AAVrg pAAV-EF1a-Flpo 87306  AAV1 AAV pEF1a-DIO-FLPo-WPRE-hGHp...testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability...

...contaminated? Well, we purposely mixed another AAV vector with AAV-68544 to validate our method. We then extracted...because of poor reagent quality. To make sure our AAV vectors are of the highest quality, we undertake a rigorous...research. To make sure that adeno-associated viral (AAV) vectors available at Addgene are suitable for in vivo...should you care about viral quality control? AAV vectors are produced by scientists - not robots. Unfortunately...DNA from the helper plasmids or the cell genome can be packaged inside the vectors during production. ...sequence of the plasmid used to create the viral prep. In a clean viral prep, like AAV-44362 shown below...cell genome, bacterial genomes, cloning vectors, and helper plasmids (Chadeuf et al., 2005, Wright et al....

...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...and share AAV Data with Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors 101: An ... and quality control are described below. AAV Production AAV distributed by Addgene has been produced ... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....determine which facility produced your viral vector lot. At Addgene, AAV particles are titered by droplet digital...

...are the elements that make an AAV plasmid an AAV plasmid.  Within the AAV genome, ITRs serve as the origin...single stranded DNA genome of AAV in the native virus. In an AAV vector plasmid, they flank the gene of interest...blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Pseudotyping Viral Vectors 101: Systemic...capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids from multiple AAV serotypes...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 gene...gene of interest in a cell or animal model. AAV plasmids contain several important elements, but among...

...efficiency of AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for...RGB AAV and red, green, or blue are the predominate colors expressed (figure 4c).   AAV Vector  Percent...efficiency of AAV-PHP.B versus AAV-PHP.eB in the CNS. A GFP transgene was packaged into either an AAV-PHP.B or...et al.   AAV Vector Percent of Neurons Transduced   Dorsal Root Ganglion (DRG) AAV.9 (parental)...Subscribe to Viral Vector Blog Posts Additional Resources on Addgene.org Search for AAV Vectors to use with...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find...much better than the AAV vectors they were derived from. Additionally, AAV.PHP.eB was used at a lower ...

...makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids...experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV:  the packaging...Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...transfer plasmid and take a look at each of its parts. Figure 1: Parts of an AAV transfer plasmid. Parts...packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector. The cloning...Transfer Plasmid video!   Now you’ve got a basic understanding of some common AAV plasmid elements! And... want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let...

...Using AAV Technologies Resources on Addgene.org Find ready to use AAV vectors at Addgene. Find AAV plasmids...during viral plasmid amplification (Fischer, KB. 2019). This means that all DIO/FLEx AAV vectors will have..., we now include guidance on the plasmid page of DIO/FLEx AAV vectors where this occurs to alert the scientist...Conclusion Using AAV vectors in your experiments requires a full understanding of vector performance in ...recombinase-dependent AAV vectors and mitigation with cross-over insensitive ATG-out vectors. Proc Natl Acad...the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When Using AAVs Three Key Considerations... cells of interest and recombinase-dependent AAV vectors for selective transgene expression. While this...

... our entire AAV inventory! Our new AAVs include: pGP-AAV-syn-FLEX-jGCaMP7b-WPRE pAAV.hSyn-FLEX.iGABASnFR... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...Recombinases Plasmid Serotype Name 50363  AAV5  AAV phSyn1(S)-DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre...indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST Chemogenetics Plasmid Serotype... of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the primary...our inventory of ready-to-use viral vectors. Here are some of the AAV we have released so far in 2019. You...Fluorescent protein and control AAV These fluorescent protein expression vectors can be used to visualize cells...

...feedback on AAVs at Addgene? Suggest a viral vector for our service; leave a comment; or take our AAV survey...released Viral Vectors 101 2nd edition — I was incredibly excited when we picked our viral vector service as...familiar with viral vectors, and especially those familiar with adeno-associated viruses (AAVs), will know just...titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both... viral vector service in 2016. We realized we could apply the lessons learned from our plasmid lab to ... high quality viral vector service, making an array of ready-to-use viral vectors available to researchers...still offer many resources for producing vectors from plasmids — including the protocols we use in our ...

... commonly used viral vectors. That’s where AAV Packaged on Request comes in.  AAV Packaged on Request ...to viral vector. To start the AAV packaged on request process, simply navigate to your plasmid of interest...AAV Packaged on Request, an expansion of our viral vectors service, has officially launched! As of today...you can request any of over 3,000 eligible AAV cargo plasmids in the Addgene repository to be packaged ...  Expanding our viral vector service Addgene has offered in-stock viral vectors since 2016, with the goal...Check our blog post on viral vector production and quality control, our AAV Packaged on Request webpage...: The AAV Packaged on Request banner.   We currently offer serotypes AAV1, AAV2, AVV5, AAV8, or AAV...

...of the AAV RepCap plasmid! Check out this blog post to learn more about the AAV transfer plasmid! References... parts of the AAV transfer plasmid  Resources on Addgene.org Learn about AAV Find AAVs for your experiment...requires some AAV.  Where do you start? Plasmids of course! You just need three plasmids to start making...making the AAVs you need for your experiment:  the packaging plasmid which contains the AAV structural ...packaging plasmid and take a look at each of its parts. Key elements of the AAV packaging plasmid The packaging... in AAV genome replication. Replication initiates from the AAV genome ITRs on the transfer plasmid, which...recombinant AAV (rAAV) is made in the lab, rep and cap are expressed from a separate plasmid in trans, ...

...our entire AAV inventory. Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! ...Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg pGP-AAV-syn-jGCaMP7b-WPRE... Viral vectors coming soon! These vectors should be packaged and available as AAV in the next few months... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources... 50457 AAV2 pAAV-hSyn-DIO-EGFP Recombinases Plasmid Serotype Name 87306 AAVrg, AAV5 AAV pEF1a-...mCherry 26973 AAVrg pAAV-hSyn-hChR2(H134R)-EYFP   Control AAV Control AAV allow researchers to not only...pGP-AAV-syn-jGCaMP7b-WPRE 104493 AAVrg pGP-AAV-syn-FLEX-jGCaMP7b-WPRE 104496 AAVrg pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE ...

...incorporated in an AAV vector (Figure 5). Efficiency and specificity of these AAV vectors for microglia was...self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand AAV (ssAAV). To ...SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia ...Dual AAV Vector System By Vaibhav Kawde To circumvent the payload limitations of traditional AAVs, the... A cis-splicing vector served as positive control (cis-ctrl). v1, vector 1; v2, vector 2; BD, binding ...Find REVeRT plasmids here! Riedmayr, L.M., et al. (2023). mRNA trans-splicing dual AAV vectors for (epi)...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  ...

...check out our latest vectors below: pGP-AAV-syn-jGCaMP7f-WPRE (104488-AAV8) pGP-AAV-syn-FLEX-jGCaMP7f-WPRE...also browse our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium...Viral vectors coming soon! These vectors should be packaged and available as AAV in the next few months. ...(121675-AAV9, 121675-AAVrg)  pENN.AAV.hSyn.Cre.WPRE.hGH (105553-AAV5) Dre vector AAV phSyn1(S)-DreO-...Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here: Cre vectors pAAV-EF1a-fDIO-Cre (121675...-WPRE (104492-AAV8) pGP-AAV-syn-jGCaMP7c variant 1513-WPRE (105321-AAV1) pGP-AAV-syn-FLEX-jGCaMP7c variant...104489-AAVrg) pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (104493-AAVrg) pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE (104495-AAVrg...

...Therapy. Existing methods for AAV quality control Historically, AAV vector QC has been sparse including...determine serotype AAV packaging is not perfect and while we think of the AAV vector genomes as containing... our viral vectors related blog posts Read our blog post on AAV titers Browse viral vectors protocols ...platform to accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed...number of AAV preparations this level of quality control is sufficient but for viral vector core facilities...demonstrated that NGS is an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing...this simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly ...

...vivo option AAV has become the preferred delivery method for in vivo CRISPR screens. AAV vectors can be made...while both AAV and lentivirus have low immunogenicity, AAV still beats lentivirus. In mice, AAV is almost...degree using different types of AAV. Expression Another advantage of AAV vectors is how the cargo — in this...workaround is to use a multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9...particularly in vivo ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in vitro, ex vivo...vivo CRISPR screens, AAV is unlikely to cause bad reactions in mice. Customization AAV offers a level of ...options, with most lentiviral vectors being packaged using VSV-G. In contrast, AAV can be packaged in a variety...

...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...Addgene.org AAV Data Hub AAV Guide AAV Packaged On Request Browse all in-stock AAV preps Browse all AAV plasmids...your choice of AAV vector for your experimental system. AAV1 AAV1 was the first viral vector approved for...Figure 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive...β3 AAV8 Laminin receptor (LR)3 AAV receptor2   AAV9 Terminal N-linked galactose3 AAV receptor2...Issa et al., 2023). AAV9 packaging plasmids are available at Addgene. AAV9 vectors are not available through...that lets AAVs “unlock” and infect specific cells. This protein shell packages the vector’s genetic material...

... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...along with AAV PHP.eB, AAV Retrograde, AAV2, AAV5, AAV8, and AAV9). Once activated, this DREADD acts through...Testing AAV inventory for the entire collection! Plasmid Serotype Name Depositor 27056 AAVrg pAAV-Ef1a-DIO...our inventory of ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released... 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 ...our control plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression...

...Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called...called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of AAV production, iodixanol...can learn more about common AAV applications by reading our Viral Vectors 101 series!  This post was originally...Assaying the Stability and Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article... addgene.org  AAV Titration by qPCR Using SYBR Green TechnologyAAV Titration by ddPCRAAV Purification ...

...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. ...