...from a lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol...Viral Vector Guides Mol Bio Protocols Introduction This protocol can be used to produce lentivirus from...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...Data Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to...efficiencies as measured by the highest proportion of GFP positive cells without limiting cell growth. Left...Left panels: bright field images; right panels: GFP channel images. Last reviewed on: August 2, 2023... Protocols Lentivirus Production Lentivirus Production You may also ...

... compound of interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your gene...generation lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty...building block Entry vectors and 5 lentiviral Destination vectors. These vectors can induce combinatorial...experiments. Gamma-retroviral vs. lentiviral vector systems Gamma-retroviral vectors have a relatively high transduction...Transfer Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase... Viral vector topic page Beginners guide to viral vectors Quick guide to all things lentivirus Resources...

...  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David Root lab is...It can be used with 2nd or 3rd generation lentiviral vectors and an envelope expressing plasmid, such ...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Sabatini’s lab is a 3rd generation lentiviral vector that contains a scrambled shRNA. This scrambled...pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)-2A-Puro (PX458), pSpCas9n(BB)-2A-GFP (PX461) expresses...pSpCas9n(BB)-2A-GFP (PX461). pSpCas9(BB)-2A-Puro (PX459) - This mammalian expression vector encodes both ...

...inventory of ready-to-use viral vectors. Here are some of the AAV and lentiviral preps we have released in ...months: Lentiviral preps for expressing SARS-CoV-2 proteins ChR2 for targeted expression DREADDs GFP dependent...plasmids into lentiviral preps to speed up COVID-19 research even further. The following lentiviral preps are...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional... activation to cells expressing both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available...pAAV-EF1a-Flp-DOG-NW Connie Cepko Viral vectors coming soon! These vectors listed below should be released at... of our viral vectors blog posts Find tips for preparing AAVs Download our viral vectors 101 eBook Resources...

...for in our reported titers. Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR...protocols. Browse our AAV vector collection. Lentivirus Production Lentiviral preparations ("-LV” catalog... 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.... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...

...Multiplexed Lentiviral Expression Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and ...expressing the Cpf1 multiplex system from a lentiviral vector.  Lentiviruses carry a (+) strand RNA copy of the...expression vectors which are often more difficult to transfect or package into viral vectors. Cas9 multiplexing...streamlined vectors which are easier to use across multiple expression platforms: transfection, lentiviral transduction...array plus a GFP tag. Four weeks post-infection, there was strong expression of Cpf1 and GFP in the targeted... 1) Most rely on transfection of more than one vector to express the gRNAs and Cas9. Co-transfections...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...

...generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector genome which...Research Lentiviral Vector Uses and Overview Resources on Addgene.org Find Lentiviral Vectors for Your...property makes the lentivirus an attractive choice for a gene transfer vector. Lentiviral vectors Although retroviruses...plan your experiment using the lentiviral vector system. Viral vectors have been increasingly popular ...primary cells. Amongst the different vectors used, lentiviral vector constructs have proven very useful ...transfer vectors The most widely used inducible lentiviral vector system is the tetracycline (Tet)-regulated ...expression vectors to target cells - one containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3)...

...efficiency. Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector or separate...separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells.... create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively achieved...Components of System Application Mammalian expression vector Promoter driving Cas9 expression can be constitutive...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression of Cas9...CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles. Transient or...

...Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells...which utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems Name... out a pair of genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9 I-CHIME Inducible knockout...knockout of a single gene pXPR_068 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 L-CHIME ...types (lineage-based) pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks...of genes sequentially pXPR_220 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME...pXPR_071, a plasmid that leads to higher titer lentivirus preparations than the first generation CHIME ...

...Service Suggestion Form. Lentivirus As some of the most popular lentiviral vectors in the repository are ...Expanding the viral vector knowledge base: You can help! We’ve been expanding our viral vector educational resources... in Your Research: A Primer for Beginners Lentiviral Vector Uses and Overview AAV: A Versatile Tool for... CRISPR and so we also have lentivirus for delivering control shRNAs, GFP, and rtTA from the Tet-on system...spending time and money producing virus from select vectors in the repository yourself, you can now order ready-to-use... production process while still ordering viral vectors. Virus production can be a frustrating process ...collection. We’ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can...

... A cis-splicing vector served as positive control (cis-ctrl). v1, vector 1; v2, vector 2; BD, binding ...SARS-CoV-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia ...within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration...incorporated in an AAV vector (Figure 5). Efficiency and specificity of these AAV vectors for microglia was...microglia The REVeRT Dual AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation ... pooled screens in human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin,...license. In this iteration, the size of the lentiviral insert was minimized to reduce recombination ...

...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown... 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...deposited to Addgene, including lentiviral and AAV expression vectors for certain variants. Klapoetke...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...

...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase) to create a final lentiviral vector containing...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...containing up to 5 different elements. Lentiviral vectors constructed with the MuLE toolbox have been used...combinations of genetic elements in a single lentiviral vector provides a powerful tool to modify mammalian...

...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification...expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some... for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay... by the virally-delivered GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...expressed GFP) that I couldn’t get a good image. You could probably also do Western blotting against GFP expression...amplification products (GFP-GFP duplex) as a control. 4. After optimizing the reaction and confirming the presence...

...infecting cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation...destabilized GFP variant fused to a 4X CDRE repeat. In addition to showing that this vector is suitable.... https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article... Ca2+ (bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (...death. Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al.,... They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison to intercalating...

...components to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro...cytoplasm and can be fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4...transcriptional activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive...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 target multiple delivery systems, such as lentivirus and MMLV gag, to specific cell types. It can ...

... Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but transient...safe for clinical applications.     Lentiviral: Lentiviral vectors infect dividing and nondividing cells...reprogramming vectors available from Addgene. Another drawback is that lentiviral vectors insert in different...so either a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming...Non-integrating iPSC delivery methods Viral Vectors Adenovirus: Adenoviral vectors infect dividing and nondividing...Tet-inducible lentiviral expression of OSKM reprogramming factors: Mouse Expression Vectors Human Expression...Excisible Medium Medium Generates transgene-free and vector-free cells Genome integration Need to sequence...

.... Each destination vector contains GFP, enabling you to select cells with high GFP expression. These cells...Multiple Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing...into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type IIS restriction...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing other...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids Research 42(19) (2014): e147...

...Tet-inducible lentiviral Gateway destination vector for gene expression GFP Varies pBABE GFP - Retroviral...expression pcDNA3 GFP LIC cloning vector - C-terminal GFP for mammalian expression...pPS808 - N-terminal GFP for yeast expression pET GFP - C-terminal GFP for... vector - Lentiviral shRNA expression Tet-pLKO-puro - Tet-inducible lentiviral shRNA...expression through lentiviral delivery pLenti CMV GFP DEST - Lentiviral Gateway destination...C elegans expression GFP Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression... bacterial expression vector (see tagged expression vectors from the SGC) pPro18...

...fusion protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3, comes...will need accessory packaging vectors to generate the virus. Bacterial vectors This system is great for overexpressing...cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope...folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your applications...shuttle vectors. You may prefer the more traditional approach of an autonomously replicating vector that ...conduct your custom experiment. Fly vectors The vast majority of fly vectors are based on p-element, random...studies involve using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following the...

...three different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to ...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...through the success of the change in expression of GFP, for the stop codon should have changed from “TGA... 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode the CD90.1 selection marker...

...CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral Lu 118029...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG... shRNA coexpression CMV Lentiviral Fuchs 26646 pCAG-Cre-IRES2-GFP Cre and GFP coexpression CAG Mammalian...-cre mCherry and Cre Lentiviral Luikart 66691 FUG-T2A-cre GFP and Cre Lentiviral Luikart 66692 pRubiC-...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector Cbh Lentiviral Eisenhoffer 132551 AAV-U6-gRNA-TnT-Cre...pCDH-CB-copGFP-T2A-iCre copGFP and iCre CB Lentiviral Oka 72257 pCDH-CB-iCre iCre CB Lentiviral Oka 73350...

...Locus. Tet-On 3G On Doyon 58245 pGLTR-X-GFP Single vector lentiviral Gateway RNAi system for conditional ...Browse Tetracycline Plasmids Tet-regulated Lentiviral RNAi Vectors Curated Plasmid Collections Background ...selection TetR On Wiederschain 41393 pCW57.1 Lentiviral vector for inducible expression for Gateway cloning...options rtTA On Root 11651 pLVUT-tTR-KRAB Lentiviral vector for inducible expression of transgene or shRNA...or rtTA None On Vogelstein 25735 pSLIK-Neo Lentiviral vector for inducible expression of a miR-shRNA; selection...third-generation rtTA On Fraser 44012 pInducer20 Lentiviral vector for shRNA expression; see article for additional...Either Lung 11662 pPRIME-TET-GFP-FF3 Lentiviral, miRNA expression (PRIME) system for application in knockdown...

...for RNAi Constitutive Lentiviral Cre-regulated Lentiviral Tet-regulated Lentiviral Constitutive Retroviral...design and delivery. Constitutive Lentiviral shRNA Lentiviral vectors for constitutive shRNA expression...system. Didier Trono Cre-regulated Lentiviral shRNA Lentiviral vectors for conditional (Cre-lox) shRNA expression...expression Tyler Jacks Tet-Regulated Lentiviral shRNA Lentiviral vectors for Tet-regulated shRNA expression...Retroviral vector for shRNA expression Bob Weinberg 10675 pMKO.1 puro GFP shRNA Negative control vector for ...Contains shRNA against GFP William Hahn 10676 pMKO.1 GFP Derivative of pMKO.1 puro with GFP instead of puromycin...Addgene; browse by constitutive lentiviral shRNA, cre-regulated lentiviral shRNA, and more. Plasmid...

...titering, testing, and providing lentiviral preps of select lentiviral vectors from the repository for you.... easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert) and the reduced titer associated... can make lentivirus production frustrating - a researcher can complete an entire lentiviral production... in lentiviral format. Finally, while we can’t currently produce custom gRNA-containing lentivirus, we...providing lentiviral preps containing highly requested gRNAs including those targeting EGFP, BRAF, and... make new lentiviral preps on your own, fear not - anytime you order ready-to-use lentivirus from Addgene...you can use along with lentiviral packaging plasmids to create your own lentivirus stocks. Our research ...

...for in our reported titers. Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR... . Browse our AAV vectors collection. AAV Data Hub Lentivirus Production Lentiviral preparations are produced... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core (Link ...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot.... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...

...17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV promoter...more details about our viral vector packaging service, visit our Viral Vector Packaging Service page . Pooled... Viral Vector Packaging Service Lentivirus Browse In-Stock Lentivirus Pooled ...Ready-to-use lentivirus available from Addgene's viral vector packaging service. Browse constructs currently... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ...available in lentiviral format. Viral...and blasticidin resistance from EFS promoter. Lentiviral backbone. Zhang Cas9 and Accessories for Activating...

... Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector for your...genome engineering from a single lentiviral vector Mammalian Lentiviral expression of Cas9, dCas9, or dCas9...Mammalian/Lentiviral BsmBI none S. pyogenes Zhang pL-CRISPR.EFS.GFP 57818 Mammalian/Lentiviral BsmBI yes...Cerulean Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA...Mammalian/Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO5.sgRNA.EFS.tRFP 57823 Mammalian/Lentiviral BsmBI... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...Mammalian/Lentiviral BbsI none S. pyogenes Puro, TagBFP Yusa lentiGuide-Puro 52963 Mammalian/Lentiviral BsmBI...

...regions Joshua Mendell 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription... Firefly hPGK Lentiviral expression of firefly luciferase Paul Khavari 109395 EFS-Vector Firefly EF1α ...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson...interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of...promoter/enhancer regions using Gateway cloning. Lentiviral. Jorge Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc...N-terminal Nanoluc fusions using Gateway cloning. Lentiviral. Mikko Taipale 12563 PCD FL0X Firefly Insertion...Firefly luciferase fusions using Gateway cloning. Lentiviral. Mikko Taipale 87077 pLenti6.2-Firefly-ccdB Firefly...

...one are in a pLVX vector background, so you can package them into lentiviral vectors if you need to. Additionally...the viral protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, all...

...Blog: Tips for Titering Your Lentiviral Preps Lentiviral Pooled Libraries All Viral Plasmids Packaging Plasmids...researcher safety. Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd...Glossary of commonly used lentiviral terms Popular Lentiviral Envelope and Packaging Plasmids One safety feature...Reiser Return to Top Popular Lentiviral Transfer Plasmids The lentiviral genome (i.e., the genetic material...generation lentiviral systems Viral production Frequently asked questions about lentivirus use Glossary...cell upon infection. When lentivirus is used for research, it is the lentiviral genome that encodes genetic...table lists lentiviral transfer plasmids that can be used with 2nd and 3rd generation lentivirus technologies...

...for vectors in the “controls” category, which includes things like viral vectors that deliver GFP. ...released Viral Vectors 101 2nd edition — I was incredibly excited when we picked our viral vector service as...titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both... high quality viral vector service, making an array of ready-to-use viral vectors available to researchers...amount of vectors available and number of requests tell only part of the story. Viral vectors are, after...offering our viral vector service because we realized that having reliable ready-to-use vectors made from plasmids...our viral vector selection process to standardize our feedback and evaluation of viral vectors. This process...

...expression vectors with various promoters suitable for in vivo expression, and/or produce vectors to make...prototrophic strains. The vectors were based on the original EasyClone vectors (Jensen et al 2013) allow...CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged for convenient...donor vectors containing mAID and either Neo or Hygro resistance markers with CRISPR/Cas vectors targeting...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable...

...high-throughput manner. They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert site.... 2018. PubMed PMID: 29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated ...cells in vivo, the non-immunogenic violet-excited GFP variant Vex. To understand the impact of this gene...

...PubMed. 3. Highly Efficient Multicistronic Lentiviral Vectorswith Peptide 2A Sequences. Ibrahimi et al. (... the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA...for each gene that is expressed, multicistronic vectors simultaneously express two or more separate proteins...blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work, and how to...get started with them. Why Use multicistronic vectors? Detecting cells that are expressing your gene,...expressing your gene of interest at high levels. Unlike vectors that express screenable or selectable markers from... the same transcript. Of course multicistronic vectors do not have to exclusively be used as a means of...

...facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in..., Myc and SV40 Nuclear Localization Signals, and GFP. B) Workflow to assess Cas9-PAGE editing with viral...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with... by pre-existing antibodies against AAV9-based vectors (and vice versa), enabling repeat dosing. In a ...

... Learn about Mammalian Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.... as RNAP II promoters is commonly employed in lentiviral and retroviral constructs and we will discuss...discuss these in a future post on viral vector parts. Promoter specificity Aside from choosing a promoter based... Can be induced by IPTG. Commonly found in pET vectors. Very tightly regulated by the lac operators. Good...-inducer fucose Weaker. Commonly found in pBAD vectors. Good for rapid regulation and low basal expression...

...Bacterial Vectors - Includes tagging with mCherry, mOrange, mCerulean pET GFP - C-terminal GFP for bacterial...bicistronic vector rather than a fusion protein vector) ZsGreen1-N1 - Mammalian Expression SGFP2 495 512 ...Expression pRSETA-PAGFP - Bacterial Expression tandem PA-GFP-KanR - Yeast Expression pFA6a-PA-GFP-kanMX6 - Yeast...Expression pFA6a-link-yEPAGFP-CaUra3 - Yeast Expression mPA-GFP-N1 - Mammalian Expression mPA-GFP-C1 - Mammalian...then scientists have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set ...mCherry LIC cloning vector (6B) - Mammalian Expression pLV-mCherry - Mammalian Lentiviral Expression pCS2+... mRFP Tsoulfas Lab Lentiviral Plasmids - Fluorescently tag your gene for lentiviral expression Verkhusha...

...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, ...tools. pLV hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific Demethylase...Core, as well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds ...Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP... targeted enhancers. Although the lab used this vector in chick embryos, it also functions in mammalian...and this fusion allows for packaging into viral vectors and targeted delivery into hard-to-manipulate cell...targeting to a wider set of genomic targets. These two vectors are part of the larger pMVP/pMAGIC cloning system...

...inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional...highlights from the past couple months: A new lentiviral CRISPR library from Alec Kimmelman targets mouse...the new selection of ready-to-use AAV9 and AAVrg vectors. New red-shifted INTRSECT 2.0 calcium sensor available...your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff/Fon-sRGECO ...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...

...- #61427 are intended for lentivirus production : 61425 : lentiviral vector encoding dCAS9-VP64 with 2A... in two formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9...activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with 2A GFP 61424 : sgRNA... 2A Blast resistance marker 61426 : lentiviral vector encoding the MS2-P65-HSF1 activator helper complex...Nature 2015). The dual vector system uses S. pyogenes Cas9 (SpCas9), using one vector to express SpCas9, ...(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH fusion ...61408 : Targeting vector for the mouse Rosa26 locus; Used to make Cas9 knockin mouse Vectors for cancer modeling...

...categories (e.g., for a lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression...the vector backbone you cloned your insert into and fill out the appropriate boxes with the vector backbone...backbone manufacturer, vector backbone size, total size of your vector + insert, any modifications you ...provide any full or partial plasmid sequences and any vector maps. The more sequence data available, the better... was a part of the insert (when cloned into the vector backbone). If your plasmid has more than one insert...you made to the backbone, and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer...enter "Feng Zhang (Addgene plasmid # 52961)".   The vector type should describe how your plasmid is intended...

...Adenovirus Guide Virus Biosafety Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook Adenoviruses are ... in E3 and insertion of GFP expression cassette. Bunz 179202 pAd5-B6/7 ∆E3-GFP Adenoviral A block for ...regions present in pAd5-B6 ∆E3-GFP and pAd5-B7 . Bunz 179203 pAd5-B1-deltaE1-GFP Adenoviral A block for the...production of GFP-trackable viruses containing transgene under CMV promoter Vogelstein 16407 pAdEasy 2-GFP beta-gal...Collection of adenovirus shuttle vectors and strains Viral...with large inserts Vogelstein 179201 pAd5-B6 ∆E3-GFP Adenoviral A block for the AdenoBuilder genome assembly...system. Block 1 with deletion in E1 and insertion of GFP expression cassette. Bunz 16402 pShuttle Shuttle ...

...engineered into viral vectors, so if you want to use them, you’ll need to use the vectors (or redesign the ...interest (analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could ...plate reader, or any other instrument which measured GFP levels and converted that signal into a readable ...Biosensors Viral Service: Biosensor AAV Viral Vectors 101 eBook Resources on the Addgene blog An Overview...ready to deliver your sensor, whether you choose lentivirus, gamma retrovirus, or AAV as your vehicle, we...

...SspB-mCherry-p60 katanin module, which are also available as lentiviral plasmids if you want to make a stable cell line...activity can be monitored by phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing...membrane-targeted SuperPLDx30 (magenta) and PA probe GFP-PASS (green). After 30 minutes of blue light illumination...Additional resources on the Addgene blog Viral Vectors 101: Optogenetics Additional resources on Addgene.org...

...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...of producing infectious virus (similar to lentiviral vectors) because they do not encode all of the necessary...on the Addgene Blog Read Other Posts on Viral Vectors Check out Our Scientist's Guide to Ebola Contribute... viral vector...non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific gene start and gene...

...MSCV-IRES-GFP MSCV For transgene expression and GFP marker Reya 21654 pMSCV PIG (Puro IRES GFP empty plasmid...γ-Retrovirus Guide Biosafety Guide Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook γ-Retrovirus (gamma-retrovirus...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...with puromycin or screen for GFP Bartel 32702 pMSCV-loxp-dsRed-loxp-eGFP-Puro-WPRE MSCV Conditional overexpression...Plasmid for transgene expression; also expresses GFP. Also see pMIG-w , a variant that has WRE, a post-transcriptional...of dsRed and the activation of your gene fused to eGFP expression Clevers 18760 MSCV IRES Luciferase MSCV...

.../ARS vector (Leu2) that contains dCas9 fused to N 51025 pSLQ1661-sgMUC4-E3(F+E) Lentiviral vector that... pU6-sgGFP-NT1 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GFP (NT1)...CRISPR imaging 51024 pSLQ1651-sgTelomere(F+E) Lentiviral vector with sgRNA targeting human telomeres 46913 ...pSLQ1658-dCas9-EGFP Human expression vector containing dCas9 that is fused to 2x NLS and EGFP for CRISPR ...anhydrotetracycline (aTc)-inducible dcas9 gene on a p15A vector with chloramphenicol resistance. The second plasmid...constitutive sgRNA expression cassette on a ColE1 vector with ampicillin resistance, wherein the N20 can...constructs 46910 pHR-SFFV-dCas9-BFP Human expression vector containing SFFV promoter, dCas9 that is fused to...

...Viral Vectors Lentivirus Oka Lab Lentiviral Vectors Baylor College of Medicine Gene Vector Core Lentiviral...series of lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors Within this...Lentiviral Vectors deposited by Kazuhiro Oka from the Baylor College of Medicine Gene Vector Core Viral...and Adenoviral Vectors You may also like... Lentiviral Guide Adenoviral Guide Viral Vector Plasmids and ...Addgene Blog: Your Lentiviral Plasmid FAQs Biosafety Resource Guide Lentiviral Transfer Vectors Adenoviral Vectors... Protocol Lentiviral Vector ID Purpose pCDH-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter ...these transfer vectors and generating infectious virus. Protocol: Lentivirus Transfer Vector Production Protocol...

...efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector or separate...separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable,...Components of System Application Mammalian expression vector Cas enzyme promoter can be constitutive (CMV, EF1alpha...typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive cells, or selection...lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or...CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles (for details...