...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...

...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson...105538 pENN.AAV.TBG.PI.ffLuciferase.RBG Firefly TGB AAV expression of firefly luciferase and GFP James ...Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Renilla EF1α AAV9-CRISPR knockout...luciferase with a V5 tag Kevin Janes 83281 pAAV-CAG-FLuc Firefly CAG AAV expression of firefly luciferase Mark...Mark Kay 105532 pAAV.CMV.ffLuciferase.SV40 Firefly CMV AAV expression of firefly luciferase James Wilson...transfection Sidi Chen 118412 ssAAV-EF1a-FLuc-WPRE-HgHpA_bac_293 Firefly EF1α AAV expression of firefly luciferase...luciferase Paul Schulze-Lefert 83282 pAAV-CAG-RLuc Renilla CAG AAV expression of renilla luciferase Mark...

...    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 ...

...pyogenes Zhang AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR 60231 Mammalian/AAV SapI none...pyogenes mCherry Kuhn AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR 60229 Mammalian/AAV SapI none S. pyogenes...pyogenes Zhang AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR 60226 Mammalian/AAV SapI none S... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...pX601 61591 Mammalian/AAV BsaI yes, cut S. aureus Zhang pX602 61593 Mammalian/AAV BsaI yes, cut S. aureus...pyogenes Joung AAV:ITR-U6-sgRNA (Backbone) PCB-FlPO-WPRE-syntetisk pA-UTR 68347 Mammalian/AAV none S. pyogenes...Cerulean Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA...

...-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 ...

...pPS808 - N-terminal GFP for yeast expression pET GFP - C-terminal GFP for...gene expression GFP Varies pBABE GFP - Retroviral gene expression pMKO.1 GFP - Retroviral ... from the SP6 promoter pCS2 mt-GFP - Create myc-tag, GFP-tag fusions for transcription ...C elegans expression GFP Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression...expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian expression...expression MSCV-IRES-GFP - Mammalian retroviral gene expression with GFP selectable ...cassette pBTG (pBigT-IRES-GFP) - pBigT modified to contain IRES-GFP Hynes Lab ...

... jGCaMP7 High-performance GFP-based calcium indicators (Constitutive or Cre-dependent) High-performance...Calcium erGAP3 (GFP-Aequorin Protein) for imaging of Ca+ dynamics in endoplasmic reticulum GFP-Aequorin Protein...FlincG3 (GFP-based cGMP sensor) for imaging in C. elegans neurons Using a Robust and Sensitive GFP-Based ...expression of GCaMP6m/s in cells depdent on Cre, Flp, or Vcre Comprehensive Dual- and Triple-Feature Intersectional...Localization Optogenetics Plasmids Viral Service: AAV Biosensors Fluorescent biosensors are proteins designed...associated with the article. We also offer ready-to-use AAV preparations of select plasmids; these are noted ...Genetically Encoded Calcium Indicators (Constitutive or Cre-dependent) jGCaMP8 Fast Genetically Encoded Calcium...

...You may also like: Viral Service: All AAV Viviana Gradinaru PHP AAV Serotypes Blog: Viral Vectors As part...actin (CBA) GFP Control Sergei Zolotukhin Citation Information AAV2(Y444F) When using the AAV2(Y444F) serotype...We provide high quality AAV preps from select plasmids in the repository. Browse the University of Florida... Viral Vector Packaging Service AAV University of Florida Serotype Testing Panel for ...viral vectors undergo quality control, including AAV titration by ddPCR, in vitro and (when possible) ...transduction of the mouse retina by tyrosine-mutant AAV serotype vectors. Mol Ther . 2009 Mar;17(3):463-71...One . Jun 8;10(6). PMID: 26052939 Lu, et al. 2016. AAV-mediated transduction and targeting of retinal bipolar...

... 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...

...genome-wide screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector...vector and used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only...typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive cells, or selection...transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich positive.../or gRNA Infects dividing and non-dividing cells AAV is the least toxic method for in vivo viral delivery...mutant allele of a gene ( point mutant )? Increase or decrease expression of a target gene? Once you have...High-fidelity Cas enzymes increase specificity. Dual-nickase approach increases specificity but is less ...

... Nat Biotechnol. 2023 May 22. See More AAV Viral Preps Find AAV viral preps for systemic delivery of viral...Noteworthy: AAV.rTH.PI.Cre.SV40 ready-to-use viral prep useful for dopamine research. CBA-driven, Cre-dependent...research. Disease Info Plasmid Collection CRISPR Tools AAV Viral Preps iPSC Differentiation Factors Antibodies...Oct 18. Target neural oxytocin receptors using an AAV-CRISPR/Cas9 strategy for gene editing across divergent...Cre-dependent expression of diphtheria toxin receptor (DTR)–GFP fusion protein. Azim et al. Nature. 2014 Apr 17. ...blood brain barrier integrity, and using CRISPR screens to understand more globally how neurons function...antibodies and scFvs for neuroscience research created with high-volume hybridoma sequencing on the NeuroMabSeq...

...pAdTrack series contains an IRES-GFP construct that enables co-expression of GFP with the transgene of interest... Adenoviral Vectors Are adenovirus and AAV different? Yes! AAV is a small single-stranded DNA parvovirus...for replication. For more information about AAV, read our AAV guide . Can I make a stable cell line with...vectors that contain IRES-GFP pShuttle Class of transfer vectors that do not contain GFP Additional Vocabulary...throughout virus production. During experiments, GFP can be used to sort cells infected with adenovirus...double-stranded DNA viruses. They are not related; however, AAV requires the presence of adenoviral genes E1, E4,...chance of recombination events creating replication-competent virus increases with each round of amplification...

...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...

...hairpin and GFP under pTREtight promoter None Either Elledge 35625 pAAV-Ptet-RFP-shR-rtTA AAV; shRNA cloning...option rtTA On Kowarz 16623 pBI-GFP Expression of your gene of interest & GFP from a bidirectional tet-responsive...Either Lung 11662 pPRIME-TET-GFP-FF3 Lentiviral, miRNA expression (PRIME) system for application in knockdown...Safe Harbor Locus. Tet-On 3G On Doyon 58245 pGLTR-X-GFP Single vector lentiviral Gateway RNAi system for ...generation; contains expression cassette for TetR-P2A-GFP; see article for additional constructs TetR On Geley...inducible promoter at its 3' end, which controls GFP expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA...16542 pBI-MCS-EGFP Expression of your gene of interest (MCS with a β-globin poly A) & EGFP from a bidirectional...

...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...

...genome-wide screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector...vector and used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit (only...typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive cells, or selection...transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich positive.../or gRNA Infects dividing and non-dividing cells AAV is the least toxic method for in vivo viral delivery...mutant allele of a gene ( point mutant )? Increase or decrease expression of a target gene? Once you have...High-fidelity Cas enzymes increase specificity. Dual-nickase approach increases specificity but is less ...