... of the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids...Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9-SSAP with...Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) by: Brian O'...developed the multiplexed version of the Cre-mediated AAV Targeted Evolution method (M-CREATE), in which one...this system could potentially be packaged into an AAV system. dCas9-SSAP will make the editing of kilobase-scaled...capsids are derived from AAV9.  Of those shown here, only AAV.CAP-B10 causes brain wide expression, with ...is true in both mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the liver is an ...

... 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...

...-mRuby3 (AAV1) pGP-AAV-syn-jGCaMP8f-WPRE (AAV Retrograde) pGP-AAV-CAG-FLEX-jGCaMP8m-WPRE (AAV Retrograde...constructs: pAAV-CAG-dLight1.2 (AAV1) pGP-AAV-syn-FLEX-Ace2N-4AA-mNeon-ST A122D WPRE (AAV1) pAAV-hSynapsin1...pAAV-CAG-SomaGCaMP6f2 (AAV9) Plus, additional AAV serotypes of the most recent jGCaMP8 calcium sensors: pAAV-hSynapsin1...Retrograde) pGP-AAV-syn-FLEX-jGCaMP8m-WPRE (AAV5)   There are hundreds of other biosensors in Addgene’s catalog...small size (~1500 bp) makes iGECInano suitable for AAV packaging.    Find iGECInano plasmids here: https...biosensors already in our collection have newly-available AAV preparations added this year. We’ve added new viral...axon-GCaMP6s-P2A-mKate2 (AAV9) pAAV-hSynapsin1-GCaMP6s-P2A-mKate2 (AAV9) pAAV-hSynapsin1-GCaMP6s-P2A-mRuby3 (AAV9) pAAV-CAG-SomaGCaMP6f2...

... enough that it can be packaged into AAV. The lab used single-AAV delivery of Nme2Cas9 and the sgRNA to...Nature Find the SECURE base editors All-in-one AAV delivery of Nme2Cas9 with gRNA cassette Erik Sontheimer...in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR interference in Candida albicans...

...developed another AAV toolkit - this time using the Staphylococcus aureus Cas9 (SaCas9). This AAV-based genome...base editors Spatially and temporally regulated AAV-based genome editing In 2016, Jonathan Ploski’s lab...in Frontiers in Molecular Neuroscience Find the AAV-based SaCas9 genome editing plasmids String assembly...

...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...pEF1a-DIO-FLPo-WPRE-hGHpA, which is now also available in AAV5 and AAVrg. Find these AAVs at Addgene New controls, biosensor...plasmids at Addgene! CRISPR RNP electroporation and AAV donor infection (CRISPR-READI) enables genome engineering...transduction of vesicular brain cells. Find this AAV at Addgene New serotypes for the Cre-dependent ...biosensor, chemogenetic and optogenetic AAV for expression in forebrain GABA-ergic interneurons. These include...hDlx-GiDREADD-dTomato and mDlx-ChR2-mCherry. Find these AAVs at Addgene ...repressor are introduced into the hPSC genome using AAV1 and contain an EGFP reporter. Expression of the ...

... precludes delivery by adeno-associated vectors (AAV). This issue was recently circumvented by the Yang... inducible Cre recombinases can be introduced by AAV or by mating with Cre-expressing mice. Before using...screened for activity in vitro and then packaged into AAV for in vivo delivery. Multiplex gene targeting is...is also possible and is only constrained by AAV packaging limits (i.e., how many sgRNA expression cassettes...Cre-dependent expression of the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive tool...individual neuronal populations.  The Yang Lab’s AAV plasmids used to express Cre under neuron-specific...lentivirus, which has a larger packaging capacity than AAV. Such an approach was described recently by the Day...

...Introduction to AAV Find AAV Calcium Biosensor Vectors at Addgene ... tissue-specific targeting. By employing various AAV serotypes, researchers can precisely direct the calcium... ready to go. Pro tip: Vectors with ready-to-use AAV preparations are labeled with the green virus particle...3):1057-65. Resources on Addgene.org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’... aav...

...easier to deliver in low-capacity vectors like AAV. AAV (adeno-associated viral vectors) are commonly ...capacity of ~4.5 kb. At 4.1 kb, SpCas9 barely fits into AAV when you add in a gRNA and regulatory elements. To...has previously used SaCas9 (1053 aa/3.2 kb) with AAV vectors for efficient postnatal editing. LbCpf1 and...than SpCas9. These nucleases should fit well into AAV vectors, especially given that Cpf1 gRNAs are also...potential to eclipse any of the CRISPR nucleases in AAV-based applications. Get More Cpf1 Background Here...

... enhanced brain potency of AAV-PHP.eB.    Figure 4: AAV9-X1 and AAV9-X1.1 efficiently transduce... variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific AAV delivery Recombinant...an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene delivery...cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when AAV-PHP.eB was injected three weeks...-01419-x. PMID: 37430038. Endothelial-specific AAV delivery By Cythina Arokiaraj (Gradinaru lab) Another...delivery to and across brain vasculature of systemic AAVs with endothelial-specific tropism in rodents and...peptide region from variable region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial...

...interneurons with AAV-S5E2-GCaMP6f are now available as a ready-to-use viral prep in serotype AAV1. Distinguish...lab are now available as ready-to-use AAV vectors in serotype AAVrg.   ...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...Distinguish dendritic neurites from axonal neurites with AAV vectors expressing cytoplasmic targeted tdTomato ...pre-synaptic targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE from Hongkui...of GCaMP8 vectors in serotypes AAV1 and AAV5. AAV9 preps and more AAV1 versions will be released soon....

...the promoter in the AAV construct. Stereotaxic surgery  As mentioned above, AAV opsin constructs are...is local injection of an adeno-associated viral (AAV) construct using stereotaxic surgery (see videos .... The Deisseroth lab has developed many of these AAV constructs with various promoters and other genetic...However, the length of promoter that can be used in an AAV construct is limited to only a few kb so attaining...found on youtube [above] and jove). Delivery of an AAV construct using stereotaxic surgery is a neuroscience...optogenetics is dependent on successful delivery of the AAV opsin construct, proficiency in stereotaxic surgeries...are used as a critical control. Depending on the AAV serotype, promoter of choice, and size of the target...

...sighted readers.    Figure 1: An image from AAV Purification by Iodixanol Gradient Ultracentrifugation...image in Figure 1 appears on our protocol page for AAV Purification by Iodixanol Gradient Ultracentrifugation...vectors for the first time and learning how to purify AAV.  Second, you’ll identify your purpose in using that...the technical details of collecting the purified AAV–that the interfaces between layers are blurred after...are captured at the 40% layer. To collect purified AAV, insert a syringe at the interface of the 40% layer...

...specifically AAV, are also being exploited as delivery mechanisms for gene therapy. The tractability of AAV, low...of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided on an application, our...00487-6 Additional Resources on the Addgene Blog AAV: A Versatile Viral tool for Gene Expression in Mammals...Listen to Connie Cepko Discuss Gene Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand...

...and adeno-associated virus (AAV). The least risky of these four viruses is AAV – it is not known to cause...integrate at chromosome 19q13.4 qtr. (AAVS1)). For these reasons, AAV is usually classified as a BSL-1 safety...safety level. Adenovirus is similar to AAV in that it does not integrate into the host’s genome. However...bleach (plastic) or ethanol (other surfaces). For AAVs, which are resistant to ethanol and isopropyl alcohol...

...Cpf1 multiplexing: Transfection, lentivirus, and AAV Cpf1’s ability to process its own pre-crRNA arrays...platforms: transfection, lentiviral transduction, and AAV transduction.  In most of Zetsche et al’s experiments...array that can be processed by Cpf1. Lastly, an AAV vector was used to multiplex edit 3-4 genes in a ...experiments, cells were infected with a 1:1 ratio of two AAVs,  one expressing Cpf1 and the other a crRNA array...Experiment A Match Made in Heaven: CRISPR/Cas9 and AAV Resources on Addgene.org Browse Our CRISPR Multiplexing...

... it is too large to be effectively packaged into AAV particles for delivery into mammalian cells, especially...archaea are much smaller and would be ideal for such AAV packaging, but their activity in mammalian systems... Cas12a based systems, making it well suited for AAV packaging. Most importantly, this CasMINI system ...effective at genome editing when delivered in an AAV system, and demonstrated that the new system was ...methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in Nature Chemical Biology Find...

...optogenetic experiments. Intracranial injections of AAV are commonly used to deliver channelrhodopsins to...collected following intracranial injection of an AAV expressing a ChRger. In both experiments, neurons...Designer channelrhodopsins paired with a systemic AAV allow for minimally-invasive optogenetics While ChRgers...delivered ChRgers by intravenous injection of PHP.eB, an AAV capsid capable of broad transduction of the nervous... on the Addgene blog Read more about the PHP.eB AAV capsid Read more about optogenetics Learn about the...

...also rolled out an updated AAV Data Hub to share curated information about AAV usage in research applications...applications. Scientists around the world can upload AAV validation data, sharing what works and, equally importantly...

...service provides ready-to-use AAV and lentiviral preps! Download the green AAV background.       Some of...the image background.     If you’re working with AAV or lentivirus in your research, you might want to... and needed a break so they're hanging out with Aavery in this Zoom video background. Want the purple ...bacteria video background with mascots Blugene and Aavery? Download the 720p version or download the 1080p...