We narrowed to 84 results for: aav vectors expressing gfp
-
TypeBlog Post...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)...
-
New and Upcoming Viral Vectors - May 2020
TypeBlog Post...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... -
New and Upcoming Viral Vectors - Spring 2019
TypeBlog Post... 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... -
New and Upcoming Viral Vectors - June 2019
TypeBlog Post...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... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog Post...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... -
Viral Production at Addgene
TypeBlog Post... 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... -
Multiplex Genome Editing with CRISPR-Cpf1
TypeBlog Post...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... -
New Viral Vectors - Fall 2024
TypeBlog Post...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... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog Post... 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(... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog Post..., “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. ... -
Hot Plasmids Spring 2024
TypeBlog Post...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... -
CRISPR 101: Mammalian Expression Systems and Delivery Methods
TypeBlog Post...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... -
New and Upcoming Viral Vectors - September 2019
TypeBlog Post...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... -
Neuronal labeling with Spaghetti Monster
TypeBlog Post...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 ... -
Viral Vectors 101: Biosensors
TypeBlog Post... 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... -
Viral Vectors 101: Parts of the AAV Transfer Plasmid
TypeBlog Post...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... -
Hot Plasmids and Viral Preps - May 2021
TypeBlog Post...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... -
Using Addgene's New Viral Service for Your Research
TypeBlog Post...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... -
Plasmids 101: FLEx Vectors
TypeBlog Post...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... -
Hot Plasmids - August 2020
TypeBlog Post...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... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog Post...systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial expression vector, use with IPTG induction... expression. CAG-FLEX: CAG promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin...neuronal expression. hSynap-FLEX: Synapsin-1 promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP...transmembrane domain in the mammalian expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol...induction to express soluble protein CAG: Generic strong promoter. Formulated as AAV-DJ for neuronal expression...FLP-dependent expression GFAP: Glial fibrillary acidic protein (promoter). Good for Glial expression (not neurons...lab swapped eGFP for sfGFP to create SF-iGluSnFR. This improved variant has higher expression levels in ... -
Popular Retroviral Vectors and Their Uses in Scientific Research
TypeBlog Post...Transfer Vectors Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...cloning and expressing your gene of interest with GFP expression. Cloning and expressing RNAi pMKO.1 ... handling and expression of these vectors when used in experiments. Retroviral vectors make up a large...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...interest. Lentiviral vectors Popular Lentiviral Transfer Vectors Expressing your gene of interest ...lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty backbones...building block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...deposited to Addgene, including lentiviral and AAV expression vectors for certain variants. Klapoetke et al... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum...strain in which Cre expression causes the induction of both DsRed fluorescence and GFP shRNA. Mice carrying...carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown in eyes and clonogenic... -
Viral Vectors 101: Calcium Sensors
TypeBlog Post...packaged into viral vectors and ready to go. Pro tip: Vectors with ready-to-use AAV preparations are labeled...blog Viral Vectors 101: Biosensors Viral Vectors 101: Viral Applications Viral Vectors 101: An Introduction...Introduction to AAV Find AAV Calcium Biosensor Vectors at Addgene ... for neuron-specific expression (hSyn promoter) and pan-mammalian cell expression (CAG promoter). An improved...mammalian cell expression, and compatible with constitutive or Cre-dependent expression. These sensors...jGCaMP7 and jGCaMP8. Viral vectors So, what do viral vectors have to do with all of these cool ...ones? Many of the available biosensors use a single GFP or CFP/YFP-derived FRET pair. If you need to free... -
Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation
TypeBlog Post... of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...guide to viral vectors Find all of our viral vector blog posts Download our viral vectors 101 eBook Additional...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes...improve their AAV yields. Given the wide range of downstream applications scientists rely on our AAV preparations...adapted by vector cores and laboratories alike as an easy and inexpensive method to improve AAV yields. ...pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering...been actively investigating ways to improve viral vector production. Whether redesigning viral packaging... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog Post...transduced at levels similar to that of a single AAV expressing GFP. The lab tested the base editors in vivo ...here. AAV delivery of split base editors Article contributed by Jennifer Tsang AAVs are a great ...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi....voltage indicators, however, fall short compared to GFP-based sensors in terms of response and brightness...parts, selectable markers, transformation acceptor vectors, and sequences for genomic integration by homology...use. This kit is designed for use with acceptor vectors from the MoClo Toolkit from Sylvestre Marillonnet...increases temporal and spatial precision of gene expression and reduces toxicity as compared to previous ... -
General Transfection
TypeProtocol...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...polyethylenimine. Transfections allow for transient expression of a gene of interest in a target cell line and...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X 293T cells were ...here by the amount of green fluorescent protein expression (green in the right panels) with a limited effect...of HEK293T cells optimized for viral production (AAVpro or Lenti-X), but any HEK293T cell line should work... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog Post...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...seen through the success of the change in expression of GFP, for the stop codon should have changed from...available including: pAAV-hsyn-GRAB_DA2h in AAV9 New GRAB-DA and 5HT sensors New AAV tools to selectively...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...from the Dimidschstein Lab More GCaMP8 ready-to-use AAV preps ... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog Post...together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this...not increased compared to a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs mediate genome...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...co-infection of two AAVs. Cas9 orthologs: shorter, but just as potent and specific? The two AAV strategies described...for in vivo studies. Testing AAV-SaCas9 in vivo To test the efficiency of AAV-SaCas9 in vivo, the Zhang ...proteins with AAV, at just 1,108 and 986 amino acids, respectively, the size of both is within AAV’s packaging... -
Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative
TypeBlog Post...destabilized GFP variant fused to a 4X CDRE repeat. In addition to showing that this vector is suitable...cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation sequencing...manipulations such as injection of precise titrations of AAV or heat-shocking the organism. The SPARC system uses...the stop cassette is removed, the effector is expressed. By truncating one of the recombination sites .... https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article...calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin is required for virulence... Ca2+ (bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (... -
Hot Plasmids February 2024
TypeBlog Post...an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene delivery...from variable region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific...Importantly, AAV1-X1 is not neutralized by pre-existing antibodies against AAV9-based vectors (and vice ... 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...cultured human neurons, with broader neuronal expression than AAV9. This new capsid will be a valuable tool...cells in mice lacking LY6A using AAV1-X1 (Chen et al., 2023); then, when AAV-PHP.eB was injected three weeks... -
Hot Plasmids: Spring 2025
TypeBlog Post...therapies. Using AAV vectors with enhancers that drive cell-type-specific expression has been a promising...backbones special: Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible...proteins, and stay tuned for mammalian expression and AAV plasmid sets launching later this year! Find Addgene's...viral preps: Enhancer-AAVs targeting interneurons By Mike Lacy Targeting gene expression to specific cell ...colleagues characterized a series of new enhancer-AAV vectors targeting specific classes of cortical and striatal...Enhancer-based targeting of PV interneurons in mice. A) AAV expression of dTomato reporter (magenta) under the control... share our very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed right here at... -
Hot Plasmids - November 2023
TypeBlog Post... ideal for size-limited AAV delivery. PE6c and PE6d are optimized for dual-AAV systems. These variants...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters...transcriptional activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element...to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications...fluorescent signal in the ticker tape from cFos::GFP-iPAK4 within an hour. Analyzing the ticker tape images...to red at 12 h, and PMA activation induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted...to create a three-dimensional atlas of protein expression in the adult mouse brain using thick-section ... -
Hot Plasmids: Summer 2024
TypeBlog Post...2024). In MagIC-cryo-EM, target proteins (expressed as a GFP fusion in this proof-of-concept study) are...capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected...than SpyCas9, allowing them to be packaged in AAV vectors, potentially with multiplexed targets. .... The team showed that AAV-delivered CHARM could silence the expression of prion protein in mouse ...SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines ...) 3D model of the obtained structure of the H1.8-GFP-bound nucleosome. Images adapted from Arimura et ...gene product; it simply turns the target protein expression off. Plus, DNA methylation is a long-lasting ... -
Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options
TypeBlog Post...of interest. In our studies, we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or...an aid, in the expression of the protein of interest. The green fluorescent protein (GFP) and mCherry, ...successful expression of the protein of interest, an alternative could be bicistronic expression of the fluorescent...the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other...proteins provide a versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore...’ tags to identify proteins Given the limited expression of hM4Di‐mCherry at the plasma membrane in monkey...years ago (Field et al., 1988). Thus, we injected AAVs carrying hM4Di-HA in the brains of mice and monkeys... -
New and Upcoming Viral Vectors - December 2019
TypeBlog Post...our entire AAV inventory. Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! ... 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...104489 AAVrg pGP-AAV-syn-jGCaMP7b-WPRE 104493 AAVrg pGP-AAV-syn-FLEX-jGCaMP7b-WPRE 104496 AAVrg pGP-...pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE 104495 AAVrg GP-AAV-CAG-FLEX-jGCaMP7s-WPRE 105714 AAV8 pAAV-Ef1a-fDIO-GCaMP6s...our inventory of ready-to-use viral vectors. Here are some of the AAV we have released in the last few months...Control AAV allow researchers to not only interpret their results, but also confirm expression in target... -
New Viral Vectors - March 2024
TypeBlog Post...pAAV.CMV.PI.EGFP.WPRE.bGH AAV PHP.eB Controls Wilson New viral vector pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls...as seven vectors newly available in the AAV PHP.eB serotype. We've also added two viral vectors that can... serotype pGP-AAV-CAG-FLEX-jGCaMP7s-WPRE AAV PHP.eB Biosensor Kim New serotype AAV-mDlx-jGCaMP8s-WPRE... New viral vector pAAV-hSyn-JEDI-2P-WPRE AAV9 Biosensor St-Pierre New viral vector pAAV-CAG-RdLight1...Notes AAV-CAG-jGCaMP8s-WPRE AAV1 Biosensor Looger New serotype AAV-CamKIIa-jGCaMP8m-WPRE AAV5 Biosensor...viral vector pAAV-nEF-Con/Foff DREADD Gi-mCherry AAV8 Chemogenetics Deisseroth New viral vector pAAV.CMV.PI.EGFP.WPRE.bGH...serotype pAAV-CaMKIIa-mCherry AAV PHP.eB Controls Deisseroth New serotype pAAV-Ef1a-mCherry AAV PHP.eB... -
Viral Vectors 101: Virus Safety
TypeBlog Post...researcher to come into contact with than GFP. Similarly, if the viral vector carries an shRNA to a gene essential...and adeno-associated virus (AAV). The least risky of these four viruses is AAV – it is not known to cause... on the Addgene blog Viral Vectors 101: Types of Viruses Viral Vector Elements Tips for Preventing...integrate at chromosome 19q13.4 qtr. (AAVS1)). For these reasons, AAV is usually classified as a BSL-1 safety...cellular function, this would also be a higher risk vector. While it is unlikely that you, or anyone else ...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... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...blog Viral Vectors 101: AAV Variables That Matter Viral Vectors 101: An Introduction to AAV References...recall, are the outer shell of an AAV vector, which determine the vector’s tropism, including its ability.... Selection of an Efficient AAV Vector for Robust CNS Transgene Expression. Molecular Therapy - Methods...payloads. Since most systemic vectors are primarily derived from AAV9, the vectors have to be administered ... For example, AAV-PHP.B doesn’t lead to enhanced transgene expression, compared to AAV9, in BALB/cJ mice... of target gene expression (EGFP, in the depositing manuscript) across the brain. AAV-BI-28 has been tested... against AAV9-based vectors (and vice versa), enabling repeat dosing (Chen et al., 2023). AAV-BI30, developed... -
Quick Guide to All Things Lentivirus
TypeBlog Post...(tTA, also expressed by your transfer vector) binds to the TRE and activates the expression of your gene...distinct expression vectors to target cells - one containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3...your experiment using the lentiviral vector system. Viral vectors have been increasingly popular in fundamental...primary cells. Amongst the different vectors used, lentiviral vector constructs have proven very useful ...fluorescent sensors. Retroviral vector background Until the development of viral vectors, gene transfer into mammalian...un-transfectable cells, labs use viral vectors and particularly retroviral vectors because they target a broader...attractive choice for a gene transfer vector. Lentiviral vectors Although retroviruses are still in use...