...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase... and flexible system can be used to construct complex lentiviruses for modification of mammalian cells... to all things lentivirus Resources on Addgene.org Gamma-retrovirus Guide Lentivirus Guide ...to choose from, for example gamma-retrovirus, lentivirus, Adenovirus, and Adeno-associated virus (AAV)...family, which includes the gamma-retrovirus and lentivirus genera, have the unique ability to integrate ...plasmids are listed here. Please note that both lentiviruses and gamma-retroviruses use the same gene products...Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning and...

... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma Our 293T...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...production protocols, titering methods, and quality control are described below. AAV Production AAV distributed...probes targeting the ITR elements and an internal control of known titer (protocol modified from Lock et ... April 2016) also performed by ddPCR.  Quality Control Addgene ensures high quality viral vectors by optimizing...production protocols and performing rigorous quality control (QC). The specific QC experiments performed varies...

...library expresses EGFP for easy visualization via direct fluorescence. Morris Control Lentivirus Browse our ...Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector, CMV...-to-use lentivirus available from Addgene's viral vector packaging service. Browse constructs currently... Viral Vector Packaging Service Lentivirus Browse In-Stock Lentivirus Pooled CRISPR Libraries Cas9 Pooled...Pooled Barcoding Libraries Control Addgene's lentiviruses are made from plasmids that are currently in ...along with 1000 non-targeting controls Puromycin Doench , Root Cas9 Lentivirus Cas9 viruses can be used to... 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 ...

...Production Titer Quality Control Resources Lentivirus Production Titer Quality Control Resources Overview of...'s viral production methods and quality control for lentivirus and AAV. Viral... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma The 293T...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...production protocols, titering methods, and quality control are described below. AAV Production AAV distributed...probes targeting the ITR elements and an internal control of known titer (protocol modified from Lock et ...

...virus and its infectivity in check. Engineering controls, safety systems that are built into how something...functions, were designed for this purpose. One such control is dividing up the essential components of a virus...integrated by the virus. If an oncogene overexpression construct is being delivered, that is a higher risk cargo... for a researcher to come into contact with than GFP. Similarly, if the viral vector carries an shRNA ...viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV)...humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma retrovirus require...and required in BSL-2 labs. When working with lentivirus, many organizations require or recommend disposable...

...pMKO.1 puro GFP shRNA Negative control vector for pMKO.1 puro; Contains shRNA against GFP William Hahn... digest) David Root 10879 pLKO.1 ‐ TRC control Negative control vector containing non-hairpin insert David...Hahn 10676 pMKO.1 GFP Derivative of pMKO.1 puro with GFP instead of puromycin resistance gene William Hahn...11578 pSico Cre addition causes EGFP to be recombined out of the construct, activating shRNA expression ...the construct, activating shRNA expression Tyler Jacks 11579 pSicoR Cre addition causes both EGFP and ...David Root 1864 pLKO.1 ‐ scrambled shRNA Negative control vector containing scrambled shRNA David Sabatini...and shRNA to be recombined out of the construct, turning OFF shRNA expression Tyler Jacks 12084 pSicoR PGK...

...Research Order Ready-to-Use Lentivirus Browse Our Lentivirus Guide Pages  ...containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA...the different vectors used, lentiviral vector constructs have proven very useful due to their ability ...create a better, safer system. Unlike the MoMLV, lentiviruses, a separate genus of the Retroviridae family...is not restricted to in vitro cell culture as lentivirus-derived vectors are capable of transducing certain...macrophages and microglia. This property makes the lentivirus an attractive choice for a gene transfer vector... for the generation of replication competent lentiviruses (RCL) especially if you are working on HIV positive...

...hUbC-driven EGFP; can be used for cDNA expression Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and shRNA ...Plasmids Lentiviral Plasmids Additional Resources: Lentivirus Guide Addgene's Viral Service Virus Biosafety...Plasmids Packaging Plasmids Transfer Plasmids Lentiviruses, a subset of retroviruses, are some of the most...and useful types of viruses used in research. Lentivirus can transduce both dividing and non-dividing ... considerations to evaluate when working with lentivirus: these viruses are based on HIV-1, which may ...Viral production Frequently asked questions about lentivirus use Glossary of commonly used lentiviral terms... and Packaging Plasmids One safety feature of lentivirus used in research is that the components necessary...

..., or a promoterless construct. Pre-constructed Entry vectors containing Cas9, EGFP, mCherry, iRFP, tdTomato...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...of inducible promoter constructs. Up to 34 building blocks were used to construct 20kb long inserts. The...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     ...extracts (Gagoski et al 2015). He has also constructed a library of eGFP-tagged human ORF clones to allow testing... performed through the N-terminal eGFP tag carried on all constructs.   pXPG: An alternative to the...

...GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used...140 genes, including constructs with unique tissue-specific promoters expressing GFP, tetracycline response... under the control of the cI857 temperature-sensitive repressor. The temperature-controlled expression...Howarth lab created the MBPx-SpyCatcher construct. This construct is used to produce a maltose binding protein...However, the laborious construction of repair templates and guide RNA constructs, as well as the difficult... Human Kinase Domain Constructs Kit, a library of human kinase domain constructs developed through a large-scale..., Hess et al. successfully evolved wild type GFP to EGFP using CRISPR-X and subsequent FACS sorting. They...

...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...assembled into a final transgenesis construct. Plasmids constructed using Golden GATEway cloning have been...identified a family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on...and demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the...upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery and providing...enabled scientists to engineer a wide variety of GFPs with diversity in fluorescence brightness, intensity...

...comes either with or without mCherry and GFP fluorescent controls. Alternatively, you can order individual...with each cell receiving only 1 TF expression construct. Cells were cultured following a differentiation...streamline identification. While the Zhang lab controlled their multiplicity of infection so that a single...disease depends on both genetic and expression control systems to mimic illness on a molecular level. ...Resources on the Addgene Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral...

... most of these constructs, catalytically dead modifiers are also available as controls. For an up-to-date...Epigenetic modifications are an additional layer of control over gene expression that go beyond genomic sequence...user-specified locus. Researchers showed that TALE-TET1 constructs, which fused a TALEN to the Tet1 demethylase ...Gersbach’s lab has deposited mammalian expression constructs including pcDNA-dCas9-p300 Core and pcDNA3.3-...Jaenisch’s lab in plasmid form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines...pCMV-dCas9-D3A) and Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a ...with the former also available as ready-to-use lentivirus. Figure 3: dCas9-DNMT3A methylates cytosines...

...currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies such...increased activation of HIV promoter-luciferase constructs. In multiple HIV-1 latent T cell lines, as well...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 ...infection period rather than the ART-induced viral control found in most HIV-1 patients. Obstacles for CRISPR...

...more gRNAs Two separate lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs ...little enrichment of DNA targeted by a negative control gRNA or predicted gRNA off-target sites. Long-range...  A second application for CAPTURE 2.0 is the construction of spatial and hierarchical chromatin interaction...

...need to generate lentivirus containing the entire library (Figure 8D). This lentivirus will then be used...-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages...LmoCas9 and SpCas9. Acr proteins can be used to control both when and where genome editing happens. CASANOVA...Dominik Niopek’s lab , is a way to optogenetically control genome editing by fusing the anti-CRISPR protein...photosensitive LOV2 domain . Several labs have also controlled Acr activity with endogenous cell-type specific... the HDR pathway can also help. Additionally, constructs with Cas9 fused to proteins that modulate HDR...delivery to cells that are poorly transduced by lentivirus. Non-mammalian CRISPR libraries are also available...

...need to generate lentivirus containing the entire library (Figure 8D). This lentivirus will then be used...-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages...LmoCas9 and SpCas9. Acr proteins can be used to control both when and where genome editing happens. CASANOVA...Dominik Niopek’s lab , is a way to optogenetically control genome editing by fusing the anti-CRISPR protein...photosensitive LOV2 domain . Several labs have also controlled Acr activity with endogenous cell-type specific... the HDR pathway can also help. Additionally, constructs with Cas9 fused to proteins that modulate HDR...delivery to cells that are poorly transduced by lentivirus. Non-mammalian CRISPR libraries are also available...