...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...-based AAV Targeted Evolution. In this technique, a pool of mutant AAV capsid (cap) expressing plasmids...deliverable AAVs? Typically AAVs are directly injected into the brain, but this tends to limit expression to near...labeling is to inject a mix of AAV vectors that constitutively express a red, green, or blue (RGB) fluorescent...et al.   AAV Vector Percent of Neurons Transduced   Dorsal Root Ganglion (DRG) AAV.9 (parental)...

...Lentiviral preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent Cre and...restrict gene activation to cells expressing both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), ... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...also offered as AAV1 (along with AAV PHP.eB, AAV Retrograde, AAV2, AAV5, AAV8, and AAV9). Once activated...our inventory of ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released...preps SARS-CoV-2 protein expression The Krogan lab quickly shared plasmids expressing SARS-CoV-2 proteins ...2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV...

... our entire AAV inventory! Our new AAVs include: pGP-AAV-syn-FLEX-jGCaMP7b-WPRE pAAV.hSyn-FLEX.iGABASnFR...Fluorescent protein and control AAV These fluorescent protein expression vectors can be used to visualize cells...restricts expression to GABAergic interneurons. These vectors can also be used as the control vectors for experiments... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...pAAV.hSyn-FLEX.iGABASnFR pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2]...constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2])... of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the primary...

...offer a Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid...testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability...can also browse our entire AAV inventory. Our new AAVs include: Control AAVs for serotype testing jGCaMP7c... add Flp-expression AAV to our recombinase AAV inventory. Flp is another useful recombinase that can be... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...55637  AAVrg pAAV-EF1a-Flpo 87306  AAV1 AAV pEF1a-DIO-FLPo-WPRE-hGHp Other new viral vectors New optogenetics...

...CatCH-GFP reporter. Expression of GFP will only result in cells where Cre-mCherry is also expressed. If ...channelrhodopsin, fused to a fluorescent GFP marker (CatCH-GFP). The lab deposited AAV plasmids containing the active...to test for cell type-specific expression of GFP. Half (113) expressed in retinal cells four weeks post-injection...Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular cell types or...Cre-mCherry expresses in photoreceptors and horizontal cells, and Cre-dependent-GFP expresses in horizontal...horizontal cells transduced by both AAVs would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters...synthetic AAV promoters The Roska lab’s synthetic promoter AAVs allows for targeted gene expression in retinal...

... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...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...

...Lentiviral Expression Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and up to ...than one vector to express the gRNAs and Cas9. Co-transfections can lead to variable expression levels ...larger expression vectors which are often more difficult to transfect or package into viral vectors. Cas9...leading to expression of a crRNA array that can be processed by Cpf1. Lastly, an AAV vector was used to... 1:1 ratio of two AAVs,  one expressing Cpf1 and the other a crRNA array plus a GFP tag. Four weeks post-infection...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...Lab Golden Gate Assembly transfection One vector expresses Cas9 and up to 7 gRNAs Each gRNA requires...

...fold higher expression levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged ...Alias: CN3781) AAV PHP.eB Recombinases Jonathan Ting New viral prep pAAV-CAG-GFP AAV9-X1.1 Controls ...packaged in AAV9-X1.1!  We also have eight new AAV-PHP.eB vectors from the Jonathan Ting lab at the Allen Institute...carried out before. We have a bunch of other new AAV vectors as well — including next-generation biosensors...ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor  Notes AAV-GfaABC1D-Cre...Addgene's Data Hub What is a Virus (and a Viral Vector)?  AAV Variables that Matter...New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls Bryan Roth New serotype AiP13044 - pAAV-AiE0779m_3xC2...

... wanted to use an AAV vector to express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested...What AAV vector is best for my specific system?  Sharing data and community concerns AAV vectors are ...with AAV vectors in neuroscience Let’s consider my first challenge when I started working with AAV vectors...serotypes containing an expression cassette with an ubiquitous promoter to express GFP in a group of young...answers hundreds of AAV questions every year on a range of issues related to AAV vectors, many from people...evaluate how small changes in vector administration can influence AAV expression properties.  By providing...transfer to the rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(...

..., “sender” cells express GFP on the surface and “receiver” cells express an anti-GFP nanobody on the cell...with AAV-S5E2-GCaMP6f, now available as a ready-to-use viral prep in AAV1!  New ready-to-use AAV prep ...pairs. Because the anti-GFP nanobody cross-reacts with BFP, sender cells expressing surface BFP generated...response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression pattern and ...for delivering CRISPRi components by a single AAV vector are now available from Tonia Rex's lab.  vfCRISPR...are available for dual expression of cytoplasmic-tdTomato and Synaptophysin-GFP. Synapse targeting GCaMP6s...Gether lab, now available as ready-to-use AAV vectors.   ...

...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 ...found that administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled...high microglia-specific gene expression when incorporated in an AAV vector (Figure 5). Efficiency and specificity...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 ...confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters in mice. B) Quantifications...

...into an AAV transfer vector and used to generate AAV particles. Transient or stable expression of SaCas9...System Application Mammalian expression vector Promoter driving Cas9 expression can be constitutive or inducible...lentiviral transfer vector or separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and...included with each expression system as well as suitable applications. Expression System Components ...or electroporation. Transient expression of CRISPR components, expression decreases as RNA is degraded ...using cationic lipids. Transient expression of CRISPR components, expression decreases as gRNA and Cas9 protein... provide an overview of some CRISPR mammalian expression systems, the typical applications for each, and...

...also browse our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium...check out our latest vectors below: pGP-AAV-syn-jGCaMP7f-WPRE (104488-AAV8) pGP-AAV-syn-FLEX-jGCaMP7f-WPRE...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-...controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE...Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here: Cre vectors pAAV-EF1a-fDIO-Cre (121675...samples of EGFP-expressing AAV packaged in various serotypes. This collection is now complete! pAAV-CAG-GFP...

...immunogenicity, and the long-term expression of AAV delivered genes. AAV serotypes have been extensively...Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV preps ...epitope tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster...connect to each other). Combined with AAVs, FPs can be easily expressed in the brain and can be detected through... structures GFP is limited in its ability to label subcellular structures in the CNS. GFP and other FPs...resolved with smFP_FLAG than GFP.   3. Enhanced ability to distinguish weakly expressed and/or similar proteins...neuronal tracing Retrograde AAV: Making the journey from axon to nucleus AAVs CREATEd for gene delivery ...

... Biosensor AAV Viral Vectors 101 eBook Resources on the Addgene blog An Overview of AAV Encoded Calcium...analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could be a ...engineered into viral vectors, so if you want to use them, you’ll need to use the vectors (or redesign the ...platforms, such as AAV or retrovirus, can either permanently integrate and/or express long term (up to ... to achieve in whole organisms, but AAVs make it possible. In AAV systems, different serotypes can be ...plate reader, or any other instrument which measured GFP levels and converted that signal into a readable ...biosensor to specific tissues or cell (sub)types for expression and downstream monitoring. Nervous about making...

...makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids...packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector. The cloning...promoter AAVs for cell-type expression in retinal cells Resources on Addgene.org Find AAVs for your ... want to do, but it requires some AAV. You’ve never worked with AAV before, but you aren’t going to let...experiments, making AAV starts with some plasmids. You just need three plasmids to start making AAV:  the packaging... the AAV transfer plasmid and take a look at each of its parts. Figure 1: Parts of an AAV transfer...   Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...

...terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo’s lab expresses membrane tethered...interneurons. We now provide ready-to-use AAV vectors expressing GCaMP6f with this promoter. Image calcium...dendritic neurites from axonal neurites with AAV vectors expressing cytoplasmic targeted tdTomato and pre-synaptic...lab are now available as ready-to-use AAV vectors in serotype AAVrg.   ...element containing vectors developed in the Jordane Dimidschsteind lab provide expression selectively in ...several versions of GCaMP8 vectors in serotypes AAV1 and AAV5. AAV9 preps and more AAV1 versions will be released...targeted synaptophysin tagged eGFP. AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE from Hongkui Zeng's lab...

...used to activate expression of a gene under the control of the tet operator. AAV AAVs are the workhorses...Beginners Lentiviral Vector Uses and Overview AAV: A Versatile Tool for Gene Expression in Mammals Additional...have retrograde AAV that you can use to map neuronal connections. Expanding the viral vector knowledge base...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... production process while still ordering viral vectors. Virus production can be a frustrating process ...researcher Connie Cepko has been helping us with AAV testing and recently had this to say about the service...

...In the presence of Cre, GFP expression is lost, allowing for rescued expression of GeneR.   Sometimes...of Genome Engineering Using FLEx AAV Vectors? Learn about Viral Vector Elements Read Our Primer on Optogenetics...elements to turn off expression of one gene, while simultaneously turning on the expression of another! What...FLEx switch that turns BFP expression off, while turning on mCherry expression. For this FLEx switch to ...improvement of optogenetic adeno-associated virus (AAV) vectors. Listed below are a few additional uses for ...allowing for expression of GeneR. This allows a researcher to test whether or not expressing GeneR alone...Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters are delivered using AAV injection...

...optogenetic AAV for expression in forebrain GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato...pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed gRNAs...and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into ...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...