...-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...are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...small size (~1500 bp) makes iGECInano suitable for AAV packaging.    Find iGECInano plasmids here!   Matlashov...biosensors already in our collection have newly-available AAV preparations added this year. We’ve added new viral...

... from the default GFP (green) to RFP, CFP, or YFP fluorescence. In Brainbow 2.1, GFP is tagged with a ...fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...Tissue-specific Cre recombination in this mouse generates cells that randomly express either nuclear GFP, membrane...OFP (orange) (Livet et al., 2007). Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent...gene directly following the promoter is expressed. Cre-mediated deletion of one or several of these genes...system. One such system is Brainbow 2.0, which uses Cre to trigger the inversion of two genes aligned head-to-head...

...anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling...off-target effects, yet small enough to be packaged in AAV. A dCasMini-mediated adenine base editor allows for...expanded the genetic screening toolkit by optimizing a Cas12a-based method for screens. Cas12a’s ability ...underutilized in genetic screens because of its lowered editing rate. This research aimed to increase the potential...parallel. Lastly, this Cas12a-based screening method was used to create a genome-wide library capable of ...single cassette provides increased simplicity for combinatorial genetic screens, and this optimized toolkit...imaging of cellular targets Optimized genetic screening technique using Cas12a for multiplexed gene perturbation...

.... Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! Optogenetics Optogenetics...pGP-AAV-syn-jGCaMP7b-WPRE 104493 AAVrg pGP-AAV-syn-FLEX-jGCaMP7b-WPRE 104496 AAVrg pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE ...Overview of AAV Encoded Calcium Sensors). Plasmid Serotype Name 104489 AAVrg pGP-AAV-syn-jGCaMP7b-WPRE...some of the AAV we have released in the last few months. You can also browse our entire AAV inventory. ... control AAV in smaller aliquots of 20 ul for serotype testing to help you pick the best AAV for your ...26976 AAV5 pAAV-hSyn-hChR2(H134R)-mCherry 26973 AAVrg pAAV-hSyn-hChR2(H134R)-EYFP   Control AAV Control... 104495 AAVrg GP-AAV-CAG-FLEX-jGCaMP7s-WPRE 105714 AAV8 pAAV-Ef1a-fDIO-GCaMP6s Neurotransmitter ...

...binding scFv that’s fused to GFP. When the multiple copies of the scFv-GFP fusion bind the SunTag scaffold...scFv that’s fused to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based... allows for their delivery by viral vectors like AAV, which could be useful for delivering scFvs that ... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...serine with dispersed hydrophilic residues for increased solubility (Monnier et al., 2013). The linker ...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...applications where longer retention times often increase therapeutic efficacy (Ahmed et al., 2012). How...

...brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed AAV (adeno-associated virus...mouse brains expressing green fluorescent protein (GFP) (6).   The Dodt and Frank Bradke laboratories later... cranial nerve afferents labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical...addition, there is a drop in image brightness with increasing depth caused by light scattering and absorption...

...    Figure 1: AAV toolkit for targeted gene delivery to the brain. The AAV (a) capsid, (b) cargo...Systemic capsids packaging Cre-dependent transgenes can also be paired with Cre-expressing transgenic mouse...precious cargo space in AAV vectors. Dual expression systems, which use two or more AAVs to transduce the same... et al, 2019 AAV-PHP.B2 CNS Mice Yes (LY6A) Deverman et al., 2016 AAV-PHP.B3 CNS Mice...2022 AAV-PHP.eC CNS: neurons and astrocytes Mice No (LY6C1) Shay et al., 2023 AAV-PHP.V1...al., 2020 AAV-PHP.N CNS: neurons Mice Yes (LY6A) Kumar et al., 2020 AAV-PHP.B4 CNS ...2023 AAV-F CNS: neurons and astrocytes Mice No (LY6C1)   Hanlon et al., 2019 AAV-BI-28 ...

...-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked...interneurons, Gordon Fishell’s lab has developed a new AAV approach using regulatory elements that restrict ...to this cell type. To this end, they constructed AAV plasmids bearing one of the distal-less homeobox ...these tools in virtually any vertebrate. This novel AAV approach allows you to target and functionally manipulate...tool for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons here...27798629 Listen to Our Podcast Segment on This New AAV Tool    mCyRFP1: Novel fluorescent protein that...light-dependent Gal4 transcriptional activation and Cre recombination. Find PhoCl plasmids at Addgene. Zhang...

... the GFP-Puro will be inserted just upstream of a stop codon. One potential concern is if the GFP-Puro...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half, without...detailed protocol for MMEJ-mediated knock-in of a GFP-Puro cassette into a given locus, just upstream of...microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks are necessary...necessary for knock-in: one on either side of the GFP-Puro cassette and one in between the 5’ and 3’ microhomologies...microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the locus (see figure below). ... overview of PITCh. The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and 3' microhomology ...

... the supplied donor plasmid will express GFP. If there’s GFP expression, presumably the endogenous target...linked to an in-frame H2B-GFP. This visual read out allows for easy screening of different gRNAs by simply...RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using the RFP:GFP ratio...Madison lab found that H2B-GFP expressing cells exhibited a 2-4 fold increase in the number of endogenous...in the genome) and an out-of-frame H2B (histone)-GFP reporter into the genome of mammalian cells.  ... can easily and seamlessly remove the integrated GFP reporter leaving you with just your desired genomic...14 and 21 to create GEmCherry2. The Denham lab made several other small changes creating 2 other versions...

...containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA...viral titre to the number of cells transduced by a GFP encoded virus, and the reverse transcriptase activity...Viral Transductions Learn Why You Might Want to Use AAV in Your Research Lentiviral Vector Uses and Overview...lentiviral vector system. Viral vectors have been increasingly popular in fundamental and applied research ...replicate. However, Mother Nature found a way to re-create a functional virus. Indeed, by testing several ...the packaging plasmid forming a vector that could create replication competent virus. Even though this system...non-dividing cells and so researchers set out to create a better, safer system. Unlike the MoMLV, lentiviruses...

... 74307 pAAV-CMV-DIO-Magneto2.0-sNRPpA AAV 74306 pAAV-CMV-DIO-TRPV4-p2A-ferritin-sNRPpA AAV 74302 ..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...brain after delivering it via AAV injection, finding that it could increase the activity of neurons from...with unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many ...loxP-stop-loxP cassette into the Rosa26 locus, allowing for Cre-dependent expression of a GMAP inserted gene. The... of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The...plasmids, which already includes dSaCas9-VPR in an AAV transfer vector for in-vivo expression and plasmids...

...mutation can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications...amplify! Examples of this are the GC-rich ITRs of AAV genomic DNA encoding plasmids, or promoters such ...assembly decreases as the cloning complexity increases. The efficiency of recombination is increased with ... minimize false positive colonies. We routinely screen just two colonies for simple cloning procedures...

...destination vector contains GFP, enabling you to select cells with high GFP expression. These cells have...them advantageous for small capacity vectors like AAV. In addition to the mammalian option described below...plasmid contains all of the desired gRNAs. This increases the chance that all the edits you want to make... cleave outside of their recognition sequence, creating flanking overhangs. These overhangs can be customized...Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing wt humanized Cas9...the gRNAs. gRNAs are flanked by glycine tRNAs to create polycistronic glycine tRNA-gRNA (PTG) constructs...using 3 distinct spacer-repeats, you could easily create 7 unique arrays (e.g. for spacers A, B, and C, ...

...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...protein Knock-off, is made up of three components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter...these dCas9s with one of three fluorescent proteins (GFP, BFP and mCherry), the authors were able to visualize...monoclonal antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags...

...of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use AAV9 and AAVrg vectors. ...available in AAV8! Expand your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff...circularly permuted green fluorescent protein or cpGFP) to the petunia SL enzymatic receptor DAD2. SLs ...which modulates the fluorescence intensity of the cpGFP without altering that of the internal control (Fig...of biosensors could be used in high throughput screening for agrochemical compounds that have the potential...

... are currently available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and you can also find several...Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications...called JEDI-2P. Using a custom, high-throughput screening platform to apply saturating mutagenesis on 21... 1: Summary of the approach and outcomes of the screen for improved genetically-encoded voltage indicators... in-vivo imaging of voltage fluctuations. Image credit: Liu Z, et al. Cell. 2022.  Find JEDI-2p viral... Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody here! ...

...AdVs have already a packaging capacity larger than AAV vectors. This capacity is large enough to carry the... pAdSh.PGK.Cas9, pAdSh.U6.gRNAS1, and pAdSh.U6.gRNAGFP. Gonçalves says that advantages of AdVs include...templates in somatic cells of adult animals in order to create animal models for particular cancer. That’s exactly...the Memorial Sloan Kettering Cancer Center did to create a model of Eml4–Alk-driven lung cancer (Maddalo... Addgene and could be useful to labs wishing to create this kind of animal models. Start using adenoviral...guide RNA constructs, pAdSh.U6.gRNAS1 and pAdSh.U6.gRNAGFP, and also Adeno Cas9 and Adeno EA. Or if you're...Circ Res 115:488–492 . https://doi.org/10.1161/circresaha.115.304351 Holkers M, Maggio I, Henriques SFD...

...Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...This post was contributed by guest blogger Tessa Cressey. The highly pathogenic Ebola virus belongs to the...non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific gene start and gene...various viral components play in disease. Tessa Cressey is a PhD Student in the labs of Dr. Elke Mühlberger...

...in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable strain. For...process. To deposit plasmids online, you will need to create an account with Addgene before starting the process...plasmids, and adding all the plasmids at once increases the likelihood that the plasmids are given sequential...Please describe any tags or fusions (e.g. His, FLAG, EGFP, etc) on your insert. Use the drop-down menu to ...