...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...

... 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...

...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 ...

...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...

... 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...

...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...

...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...

...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...

...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...

...elements are inserted into an AAV transfer vector and used to generate AAV particles. Transient or stable...for conducting genome-wide screens using CRISPR. AAV transduction Only compatible with SaCas9 (packaging...or gRNA. Infects dividing and non-dividing cells. AAV is least toxic method for in vivo viral delivery....hard Viral delivery   Lentivirus Retrovirus AAV Cells Dividing or nondividing Dividing Dividing...

... AAV8, as well as AAV5 and AAVrg. Find the Cre-expressing viral preps here AAV-CAG-dLight1.3b from ...in serotype AAV5! Get dLight1.3b here Looking for a knock-out in your experiments? AAV-flex-taCasp3...available as a ready to use viral prep in serotype AAV5. Find AAV-flex-taCasp3-TEVp here The red-shifted channelrhodopsin...SaCas9 ABE variants. It's also one of the smallest AAV-deliverable ABE.  Find the plasmid here Jesse Zalatan...the Deisseroth viral preps here Coming soon! New AAV serotypes from the Sanford Boye lab with enhanced...include AAV1, and AAV9. Find viral preps here pAAV-hSyn-Cre-P2A-dTomato is now available in AAV1 and AAV8...Loren Looger Lab now available in AAV1. Get GFAP.SF-iGluSnFR.A184S-AAV1 here New Dual and Triple feature...

...future experiments.  Use Addgene’s AAV Data Hub. You can search the AAV Data Hub for tools to use in future...experiments. If you’ve used Addgene’s AAV tools, you can contribute data to the AAV Data Hub to help the research...

...currently uses ddPCR for AAV titrating. You can learn all about ddPCR for AAV titration and find helpful...tricks in our blog post “Droplet Digital PCR for AAV Quantification.”   Multiplex PCR Multiplex PCR, as... nucleotide concentration of starting material AAV Titrations Allelic Variations Quantifying low abundance...resources on the Addgene blog Droplet Digital PCR for AAV Quantification Plasmid cloning by PCR  Plasmids 101...