... 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...

...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...

...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 ...