... 50363  AAV5  AAV phSyn1(S)-DreO-bGHpA 121675  AAV9  pAAV-EF1a-fDIO-Cre (FLP-dependent CRE) 55634...constitutive (pAAV-Syn-Archon1-KGC-GFP-ER2) and cre-dependent version (pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2])...pAAV.hSyn-FLEX.iGABASnFR.F102G pAAV-Syn-Archon1-KGC-GFP-ER2 pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104...Our new AAVs include: pGP-AAV-syn-FLEX-jGCaMP7b-WPRE pAAV.hSyn-FLEX.iGABASnFR pAAV.hSyn-FLEX.iGABASnFR.... of jGCaMP7 in AAV1 at Addgene! See all calcium sensor AAV at Addgene. GABA sensor AAV GABA is the primary...diversified) 37825  AAV8*, AAV9*  pAAV-CAG-GFP 50465  AAV8*, AAV9*  pAAV-hSyn-EGFP *Also coming in...are some of the AAV we have released so far in 2019. You can also browse our entire AAV inventory! Our ...

...expressing both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available in AAV1. Plasmid...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional...genome-wide inhibition screening. Find the Dolcetto lentiviral prep here Control AAV Control AAV allow researchers...encoding a Cre dependent version of the DREADD hM3D is now also offered as AAV1 (along with AAV PHP.eB, ...proteins ChR2 for targeted expression DREADDs GFP dependent Cre and FLPo And more!  New lentiviral preps...offer control AAV in smaller aliquots of 20ul for serotype testing to help you pick the best AAV for your ...plasmids in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain...

...efficiency of AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for...efficiency of AAV-PHP.B versus AAV-PHP.eB in the CNS. A GFP transgene was packaged into either an AAV-PHP.B or...Search for AAV Vectors to use with your research. Visit Our AAV Guide Page for info on AAV components,...method?  CREATE stands for Cre Recombinase-based AAV Targeted Evolution. In this technique, a pool of mutant...cell-type-specific Cre, the polyA sequence is inverted only in the Cre-expressing tissues. This inversion creates a template...neurons and astrocytes. Why screen for IV deliverable AAVs? Typically AAVs are directly injected into ...PNS. A GFP transgene was packaged into either an AAV.9 or an AAV.PHP.S capsid. Both viruses were IV injected...

...channelrhodopsin, fused to a fluorescent GFP marker (CatCH-GFP). The lab deposited AAV plasmids containing the active... of Cre-mCherry and a second promoter to drive expression of a Cre-dependent fluorescent CatCH-GFP reporter...Expression of GFP will only result in cells where Cre-mCherry is also expressed. If Cre-mCherry expresses...horizontal cells transduced by both AAVs would express Cre-mCherry and GFP.   Figure 2: Synthetic promoters...Performance of synthetic promoter AAVs in mice All of these synthetic promoter AAVs were first injected into the...  Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular cell... photoreceptors and horizontal cells, and Cre-dependent-GFP expresses in horizontal cells and ganglion...

...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid...Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase AAV inventory here...testing) pAAV-CAG-GFP 50465 AAV8, 9 (20 µL size serotype testing) AAV-hSyn-EGFP Biosensor AAV The ability...Serotype Name 104496  AAV1 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE 105322  AAV1 pGP-AAV-syn-FLEX-jGCaMP7c variant...Name 119741  AAV5  AAV SYN flex PSAM4 GlyR IRES EGFP Recombinase AAV Finally, we are excited to add...some of the AAV we have released in the last few months. You can also browse our entire AAV inventory. .... Our new AAVs include: Control AAVs for serotype testing jGCaMP7c for calcium detection PSAM4 GlyR for...

...Two days later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected...Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at ... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from... and quality control are described below. AAV Production AAV distributed by Addgene has been produced ...purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV subjected...arrowhead). Scale bar = 100 nm.   AAV resources Browse AAV preps available from Addgene. After receiving...Addgene blog Find and share AAV Data with Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors...

...neuronal tracing Retrograde AAV: Making the journey from axon to nucleus AAVs CREATEd for gene delivery to the...immunogenicity, and the long-term expression of AAV delivered genes. AAV serotypes have been extensively studied... structures GFP is limited in its ability to label subcellular structures in the CNS. GFP and other FPs...Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV preps ...photostable than GFP. In addition, antibody-based amplification of their signals is weak and increases background...three decades, recombinant Adeno-associated virus (AAV) has been widely used to deliver genetic tools to...atlas enables researchers to quickly find the best AAV serotypes to target the neurons they’re studying....

...labeling Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9...-4.   Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) by: Brian...multiplexed version of the Cre-mediated AAV Targeted Evolution method (M-CREATE), in which one applies both... of the AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids...line expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target... proximity to the GFP-tagged protein (albeit slightly offset due to the nanobody-GFP interaction). The...ligase TurboID fused to a conditionally stabilized GFP-nanobody rather than a specific protein of interest...

...pAAV-EF1a-fDIO-Cre AAV5 Recombinases Esteban Engel New serotype pAAV-hSyn-Cre-P2A-dTomato AAV PHP.eB Recombinases...viral prep pAAV-CAG-GFP AAV9-X1.1 Controls Edward Boyden New serotype pAAV-hSyn-EGFP AAV PHP.eB Controls...levels compared to AAV9. You can now target these cells with AAV-CAG-GFP packaged in AAV9-X1.1!  We also ...Notes AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre...serotype AiP13778 - pAAV-AiE0450h-minBG-iCre(R297T)-BGHpA (Alias: CN3778) AAV PHP.eB Recombinases Jonathan...viral prep AiP13779 - pAAV-AiE0779m_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN3779) AAV PHP.eB Recombinases...viral prep AiP13781 - pAAV-AiE0452h-minBG-iCre(R297T)-BGHpA (Alias: CN3781) AAV PHP.eB Recombinases Jonathan...

...GBPs/GFP. The new system, CRE-DOG (Cre Dependent On GFP), is activated by GFP and derivatives GFP and ...Connie Cepko Combining Cre and GFP with Cre-DOGs The T-DDOG results showed that GFP could regulate transcription...transcription or Cre recombinase activity only in the presence of GFP. These systems, known as T-DDOG and Cre-DOG,...to bind GFP. The idea of using GFP as a scaffold suddenly seemed very realistic. Envisioning GFP as a substrate...induced in ~76% of GFP+ cells, and 100% of DsRed+ cells also expressed GFP. Using AAV constructs, Tang ... cord. Cre-DOG is also suitable for optogenetic studies, and since cells infected with AAV-Cre-DOG retain...cells cotransfected with GFP and the GBPs, GFP linked the two GBPs together to create a complete transcription...

...with AAV-S5E2-GCaMP6f, now available as a ready-to-use viral prep in AAV1!  New ready-to-use AAV prep ...sender” cells express GFP on the surface and “receiver” cells express an anti-GFP nanobody on the cell ...Synaptophysin-GFP. Synapse targeting GCaMP6s from the Rylan Larsen lab, now available in a ready to use AAV5 viral...to track cell-cell interactions By Jennifer Tsang GFP-based Touching Nexus (G-baToN) is a new tool developed...cell surface. When the cells interact, GFP is transferred to the receiver cell, and the receiver cell turns...other fluorophore/antibody pairs. Because the anti-GFP nanobody cross-reacts with BFP, sender cells expressing...Plasmids for delivering CRISPRi components by a single AAV vector are now available from Tonia Rex's lab.  vfCRISPR...

...sensor was engineered from a GFP variant and a DA receptor such that GFP fluoresces when the receptor ... Yulong Li’s lab has created the GRABDA sensor based on a similar strategy: a GFP variant bound to a DA... DA receptor, such that the GFP fluoresces upon DA-receptor binding (Sun et al., 2018). These sensors ...behaviors. Find dopamine and other biosensors in our AAV vector inventory. References Darvesh, Altaf S.,...viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources ...service Find viral protocols and videos Find biosensor AAVs for your research ...submicromolar) DA affinity and high-magnitude responses (increases in fluorescence) at low concentrations (e.g., ...

...makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases each and AAV plasmids... 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...packaged into the AAV molecule. What’s outside of the ITRs does not get packaged in the AAV vector. The cloning...ITR loss abolishing AAV packaging. To prevent ITR loss, it’s best to propagate AAV constructs in recombination-deficient...

...and Share AAV Data with Addgene's New AAV Data HubCelebrate Open Data Day with Addgene’s AAV Data Hub!...with AAV vectors in neuroscience Let’s consider my first challenge when I started working with AAV vectors... answers hundreds of AAV questions every year on a range of issues related to AAV vectors, many from people...was: What AAV vector is best for my specific system?  Sharing data and community concerns AAV vectors ...transfer to the rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(... to express GFP in a group of young rats. We found that, in this case, AAV1 (23%) and AAV6 (<5%, data ... Fig. 1: Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2 noradrenergic...

...Flp-dependent Cre AAV in AAV9 and AAVrg. See our new additions here: Cre vectors pAAV-EF1a-fDIO-Cre (121675...: pGP-AAV-syn-jGCaMP7f-WPRE (104488-AAV8) pGP-AAV-syn-FLEX-jGCaMP7f-WPRE (104492-AAV8) pGP-AAV-syn-jGCaMP7c...mCherry)-EGFP (112677-AAV5) Calcium Sensors pGP-AAV-syn-jGCaMP7b-WPRE (104489-AAVrg) pGP-AAV-syn-FLEX-jGCaMP7b-WPRE...pGP-AAV-syn-FLEX-jGCaMP7b-WPRE (104493-AAVrg) pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE (104495-AAVrg) pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE.... Our new AAVs include: EGFP-expressing AAV for serotype testing Calcium sensors and GABA sensors Optogenetics...expresses nuclear EGFP in the presence of Cre. Biosensor AAV (calcium sensors and GABA sensors) Calcium sensors... of Cre-dependent GABA sensors. See all our GABA sensor AAV inventory here. pAAV.hSynap.iGABASnFR (112159...

...self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand AAV (ssAAV). To ...Voltron2 voltage sensors AAV vectors for targeting microglia The REVeRT Dual AAV Vector System Neurodegeneration...investigators found that administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled... incorporated in an AAV vector (Figure 5). Efficiency and specificity of these AAV vectors for microglia...Dual AAV Vector System By Vaibhav Kawde To circumvent the payload limitations of traditional AAVs, the...Ace2N-mNeon-A122D and Voltron2 are available as Cre-dependent AAV1 vectors and are suitable for extended imaging...Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters in mice...

...elements are inserted into an AAV transfer vector and used to generate AAV particles. Transient or stable...common choice for conducting genome-wide screens using CRISPR. AAV transduction Only compatible with SaCas9...reporter protein such as GFP? All of these outcomes can be most effectively achieved with different CRISPR components...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells. Likely contains...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...

...al. “Cell type-specific manipulation with GFP-dependent Cre recombinase.” Nat. Neurosci. 18(9) (2015):...about Professor Cepko's recent work on developing GFP-activated proteins, read our recent blog post. Check...Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools Being Developed in the Cepko Lab...on the Addgene Blog Read our Recent Blog Post on GFP Activable Plasmid Tools Being Developed in the Cepko... with Michael Koeris Get Cepko Lab Tips on using AAVs in Your Experiments  Resources on the Addgene Website...

...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...optogenetic AAV for expression in forebrain GABA-ergic interneurons. These include mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato...vesicular brain cells. Find this AAV at Addgene New serotypes for the Cre-dependent Flpo recombinase pEF1a-DIO-FLPo-WPRE-hGHpA...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...sensors include syn-jYCaMP1s (slow variant), the Cre-dependent syn-FLEX-jYCaMP1s, and the axon-targeted...hDlx-GiDREADD-dTomato and mDlx-ChR2-mCherry. Find these AAVs at Addgene ...

...combines the Cre/lox system with optogenetics. Cre activity is controlled by red light by splitting Cre and fusing... Figure 4: The C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome... previous Cre/lox or drug-induced Cre systems (ex: tamoxifen). As the name suggests, CreLite combines ...here.        AAV delivery of split base editors Article contributed by Jennifer Tsang AAVs are a great ...transduced at levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo ... dimerize, bringing each terminus of Cre together to restore Cre activity. As soon as red light or PCB...proteins separate, inactivating Cre. This is critical for reducing Cre toxicity in long-term experiments...