...repression of your target gene, activation and repression of different genes in a single cell, and localization...Favorite Fluorescent Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts ...replication and transcription as well as reporter genes. To learn how you can use minigenomes in your research... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (... (AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are...localization of a fluorescent signal to your target gene. Read this post to find out more about how the Casilio...are widely used by the optogenetics and chemogenetics communities to activate neurons and neuronal pathways...

...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...These vectors allow you to test the function of Your Gene Of Interest (YGOI) in a controlled environment under...design. You can create a new strain with a disrupted gene by transforming wild-type yeast with an integrating...using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following the expression pattern...modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check...visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected cells... More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other...

...Versatile Tool for Gene Expression in Mammals Additional Resources on Addgene.org Overview of Viral ...pooled libraries. Whether you’d like to knockout a gene, replace it, modulate its expression, or perform...also have lentivirus for delivering control shRNAs, GFP, and rtTA from the Tet-on system. The first two can...while rtTA can be used to activate expression of a gene under the control of the tet operator. AAV AAVs ...associated virus (AAV, primarily chemogenetics tools for now but with optogenetic tools coming soon). The viral...excited that Addgene is now offering virion preparations to the scientific community. Addgene's AAV preparations...posts on the blog and pages on the Addgene website. blog.addgene.org The blog provides tips for working...

...in applying gene technology to bacteria and plants. Her research involves cloning cry genes from the soil...using a bacterial expression system such as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant... shared mission of both Seeding Labs and Addgene. The Addgene-Seeding Labs plasmid grant is a collaborative...of the Addgene-Seeding Labs Plasmid Grant awardees' projects progress. Subscribe to the Addgene blog for.... Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant Check...introduce the 5 scientists who were awarded the Addgene-Seeding Labs Plasmid grant this year. These scientists...plant biology, she coordinates the Network for the Genetic Improvement of Cowpea, a knowledge database for...

...MSCV-IRES-GFP MSCV For transgene expression and GFP marker Reya 21654 pMSCV PIG (Puro IRES GFP empty plasmid...Weinberg 10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro.... Hahn 63704 pRetroX GFP T2A Cre CMV/MSV Tetracycline inducible expression of GFP T2A Cre fusion in mammalian...) MSCV For cloning and gene expression; select with puromycin or screen for GFP Bartel 32702 pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE...your gene fused to eGFP expression Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression...Lowe 9044 pMIG MSCV Plasmid for transgene expression; also expresses GFP. Also see pMIG-w , a variant that...transfer plasmids. Want a retroviral plasmid with your gene of interest? Search our database for retroviral ...

...experiment. PITChing MMEJ as an Alternative Route for Gene Editing Michohomology-mediated end joining (MMEJ...PubMed Central PMCID: PMC5495063 Kan et al. use a GFP-BFP conversion system to assess precision genome ...template to increase efficiency of insertion and gene replacement. Care to share your favorite CRISPR ...Blugene and I represented Addgene at the recent Keystone meeting on Precision Genome Editing with Programmable...hopes to evaluate genome editing pipelines and generate a common lexicon for genome editing studies. These...desired edit, and I want to make sure that I (and Addgene) don’t forget this in the future. Here’s a list... to keep current with the latest and greatest! Addgene resources 3 Tips to Improve HDR Efficiency for ...

... (FLAG, Myc, etc.) and fluorescent protein tags (GFP, mCherry, etc.). Fluorescent tags are primarily used...Solubility Enhancer. In P. E. Vaillancourt, E. coli Gene Expression Protocols (Vol. 205, pp. 99–118). Humana...-59259-301-1:99 Resources on Addgene.org Addgene antibodies Addgene’s antibody guide Antibody protocols...having solubility issues!). Need a GST antibody? Addgene has you covered! His His tags, often called polyhistidine...Purification of His fusion proteins from E. coli is generally more straightforward and of a higher purity than...and other qualities of each tag class, they are generally best suited for their primary application. Ready...protocols  Resources on the Addgene blog Plasmids 101: Protein tags Antibodies 101: Epitope tags To Each...

...endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...during adeno-associated virus manufacturing." Human gene therapy21.10 (2010): 1251-1257. PMID: 20649475. ...have probably heard about Addgene’s viral service but did you know that Addgene also performs independent...101 eBook Additional resources on Addgene.org Learn about Addgene's viral service Learn about viral production...research? Since launching the viral service in 2016 Addgene’s research team has been actively investigating ...PMCID: PMC2957237. Additional resources of the Addgene blog Read our beginner's guide to viral vectors...

...soCoChR-GFP Syn CoChR (soma-targeted) GFP Constitutive 9 Boyden 107712 pAAV-hSynapsin-FLEX-soCoChR-GFP Syn...pAAV-hsyn-Jaws-KGC-GFP-ER2 Syn Jaws GFP Constitutive 5, 8, rg* Boyden 84445 pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] CAG...* Deisseroth 58880 pAAV-Syn-ChR2(H134R)-GFP Syn ChR2/H134R GFP Constitutive 8, rg* Boyden 100054 pAAV....Constitutive 9 Harvey 59170 pAAV-Syn-Chronos-GFP Syn Chronos GFP Constitutive 1, 5, rg* Boyden 59171 pAAV-...Constitutive 8 Deisseroth 22222 AAV-FLEX-Arch-GFP CAG Arch GFP Cre dependent 1, 9 Boyden 28305 pAAV-FLEX-...dependent 5 Boyden 99039 pAAV-CamKII-ArchT-GFP (PV2527) CaMKII ArchT GFP Constitutive 1, 5, 9, rg* Boyden 137148...EF1a/nEF Synapsin E2 regulatory element Fluorophore GFP Red-wavelength fluorescent protein Yellow-wavelength...

...Dimidschstein 135631 pAAV-S5E2-GFP-fGFP E2 regulatory element GFP, membrane-targeted GFP Control Dimidschstein ...Boyden 37825 pAAV-CAG-GFP CAG GFP Control Boyden 50469 pAAV-CaMKIIa-EGFP CamKIIa EGFP Control Roth 59462 ...104061 CAG-NLS-GFP CAG NLS-GFP Control Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Control Wilson...tdTomato Control Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden MaCPNS2 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden CAP-B10 These viral vector preparations...Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control Boyden CAP-B22 These viral vector preparations...

...multiple) rAAV-CAG::FLEX-rev:: hM4D-2a-GFP AAV1 Chemogenetics Scott Sternson New viral prep pAAV-CaMKIIa-hM3D...mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-CaMKIIa-hM4D(Gi)-mCherry AAV1 Chemogenetics Bryan ...mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-hSyn-hM4D(Gi)-mCherry AAV1 Chemogenetics Bryan Roth...AAV-PHPeB Optogenetics Jonathan Ting New viral prep AiP14033: pAAV-AiE0779m_3xC2-minBG-CoChR-EGFP-WPRE3-...AAV-PHPeB Optogenetics Jonathan Ting New viral prep AiP14035: pAAV-AiE0452h_3xC2-minBG-CoChR-EGFP-WPRE3-...pAAV-AiE0873m_3xC2-minBG-CoChR-EGFP-WPRE3-BGHpA (Alias: CN5140) AAV-PHPeB Optogenetics Jonathan Ting New viral... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...

...highlighted back in 2015, where sdAbs were used to create GFP scaffolds for transcriptional activation.  sdAbs ...from addgene.org  Plasmids encoding nanobodies at Addgene Sybody Generation Toolbox from Addgene     ...only antibodies. (A) Overview of genetic modifications made to generate the nanomouse. (VHH = variable ...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended...to human populations around the world. Here at Addgene, we love camelids too, though not just for the ...chromobodies,” which are single-domain antibodies genetically fused to fluorescent proteins and used to visualize... synthetic single-domain antibody libraries to generate new sdAbs. There has also been some interesting...

... Fig. 3: Image of Addgene’s western blot for the myc-tagged protein GFP-smFP-myc expressed from ...C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first anti-c-Myc [9E10] (...high-throughput screening platform to apply saturating mutagenesis on 21 positions in the protein sequence of the...approach and outcomes of the screen for improved genetically-encoded voltage indicators (GEVIs) based on the...kinetics. They demonstrate the use of HcKCR1 as an optogenetic silencer in experiments with mouse cortical neurons... HcKCR2, making it more suitable for in vivo optogenetic use.   Fig. 2: A) HcKCR1 is highly selective...control K+ gradients and can be used as a potent optogenetic silencer. The current work also proposes promising...

...contains circularly permuted green fluorescent protein (GFP), and nLightR uses a red fluorescent protein, mApple...Imaging Tools Resources on Addgene.org Find Biosensor Plasmids Find Optogenetics Plasmids Visit the Fluorescent... visualize their activity. In 2019, the first generation of norepinephrine sensors was developed, named...norepinephrine (Feng et al., 2019). But this first generation of tools had limited sensitivity and speed and... models of nLightG (left) and nLightR (right) generated using ColabFold (Mirdita et al., 2022). B, Representative...Speed One of the major advantages of this next generation of norepinephrine sensors is their speed. nLightG...GRABNE were more sensitive than nLightR, both generating a measurable signal after electrical stimulation...

...Fishell 83895-AAV1 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV2 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV8 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAV9 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...Fishell 83895-AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP expression in forebrain GABA-...element Gordon Fishell 83900-AAV1 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV2 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...

...Protein Purpose Principal Investigator Plasmids GFP GFP-dependent transcription factors Connie Cepko See...localization Biosensors Optogenetics Background Fluorescent proteins such as GFP have long been used for...scaffold to allow scientists to selectively modulate gene expression. Plasmid...regulate a variety of other activities. By using GFP as a scaffold, scientists can design de novo systems...modular components or take advantage of existing GFP-lines for cell-specific manipulation. Plasmids Fluorescent...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

...targeting a gene associated with diphtheria toxin lethality and were sorted into single wells to generate clonal...instance, it’s much more difficult to introduce a gene of interest into primary cells, so most primary ...Neuronal stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. ... risk of viral integration causing mutations and gene expression changes elsewhere in the genome. Second...j.cell.2015.03.028. PubMed. Find the Plasmids @Addgene: iTOP-tested proteins and reporters Previous...

...collection is now complete! pAAV-CAG-GFP (plasmid 37825) and pAAV-hSyn-EGFP (plasmid 50465) are now available...include: EGFP-expressing AAV for serotype testing Calcium sensors and GABA sensors Optogenetics Recombinases... pAAV.hSynap.iGABASnFR (112159-AAV1) Optogenetics Optogenetics continues to be a useful method for manipulating...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols... testing AAV, which are small (20 ul) samples of EGFP-expressing AAV packaged in various serotypes. This...recently released these controls: pAAV-hSyn-DIO-EGFP (50457-AAV5): The Synapsin promoter directs broad...broad, neuronal expression. AAV pCAG-FLEX-EGFP-WPRE (51502-AAV5): The CAG promoter directs ubiquitous expression...

...CAG-NLS-GFP CAG NLS-GFP Constitutive PHPeB Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Constitutive...Fluorophore/Tag Activity Serotype PI 37825 AAV-CAG-GFP CAG GFP Constitutive 1, 2, 5, 8, 9, rg*, PHPeB, CAP-B10...*, PHP.eB, PHP.S Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Constitutive 1, 2, 5, 8, 9, rg*, PHP.eB ...dependent PHP.V1 Gradinaru 83895 pAAV-hDlx-Flex-GFP-Fishell_6 Dlx GFP Cre dependent 1, 2, 8, 9, rg* Fishell 83894...PHP.eB Dimidschstein 135631 pAAV-S5E2-GFP-fGFP E2 regulatory EGFP Constitutive 1, 9, PHP.eB Dimidschstein... 9 Wilson 105598 pAAV.GfaABC1D.PI.Lck-GFP.SV40 GfaABC1D Lck-GFP Constitutive 5 Khakh 105622 pAAV.CamKII...pENN.AAV.CamKII0.4.eGFP.WPRE.rBG CamKII EGFP Constitutive 1, 5 Wilson 105535 pAAV.TBG.PI.eGFP.WPRE.bGH TBG EGFP Constitutive...

...destination vector contains GFP, enabling you to select cells with high GFP expression. These cells have...sequence between two target sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple...simultaneous multiplex editing of six different genes. This system doesn’t rely on any additional or host-dependent...from this paper at Addgene. 9. Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic ...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Browse Our CRISPR Topic...add multiple gRNAs on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll use...multiplex gRNAs, some being: Using dual nickases to generate a knockout or edit. This can help reduce off-target...