...in vitro transcription as an alternative. Based on transient double-selection of transfected cells, the...new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic... RVD domains designed for rapid TALEN assembly. Kiran Musunuru and Chad Cowan Ekker Lab TALEN Kit All ...strong nuclear localisation tags and repress transcription initiation when targeted to DNA sequences within...Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN expression is driven by the strong...reporter vector pGL4-SSA can be used for single-strand annealing assay, which enables the evaluation of...destination vectors can be used to create TALE-transcription factors and are used at the final assembly step...

...lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors Within this set of...packaging these transfer vectors and generating infectious virus. Protocol: Lentivirus Transfer Vector Production...Plasmid FAQs Biosafety Resource Guide Lentiviral Transfer Vectors Adenoviral Vectors Kazuhiro Oka and the... of lentiviral transfer vectors you can find reporter plasmids to test whether or not you've efficiently...coexpressed with tdTomato and Pac (puromycin N-acethyl-transferase) pCDH-CB-copGFP-T2A-iCre 72256 Expresses copGFP...PGK promoter Adenoviral Vectors The adenoviral transfer vectors contained within this set are derived ...Flp recombinases. Genes can be cloned into these transfer vectors using shuttle vectors pICPIS-EF1 and pICPIS-CB...

...fused to the N-terminus of LSSmOrange pLSSmOrange-C1 Orange Mammalian Express a gene of interest fused ...resonance energy transfer, or FRET , is a process by which energy is non-radiatively transferred from an excited... GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange Mammalian Express a gene of interest fused...excited donor fluorophore to an acceptor. Since the transfer of energy does not occur by emission of a photon...FRET experiments. The efficiency of the energy transfer is measured using fluorescence microscopy by exciting...fused to the C-terminus of LSSmOrange mRuby2 Red Mammalian Expresses mRuby2 (a RFP variant) commonly used ...

...Browse the Zhang lab’s comprehensive human transcription factor collection, available at Addgene. The...The collection can be used for screening transcription factors for cellular programming or cancer drug ...Library The MORF library is a comprehensive human transcription factor collection that contains 1,836 genes ...collection, can be purchased at Addgene. Each transcription factor isoform has a unique 24 bp barcode that...that simplifies identification of the transcription factor isoform in pooled screens. This collection ...collection and library can be used for screening transcription factors for applications such as cellular programming...cancer drug resistance. Protocols & References A transcription factor atlas of directed differentiation. Joung...

...protein translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery...Bacterial Ahmed Badran 209739 pAB228i24 G1 PylRS Methanogenic archaeon Bacterial Ahmed Badran 209740 pAB228q12...jannaschii Bacterial Ahmed Badran 209741 pAB228r3 Mlum1RS Bacterial Ahmed Badran 209742 pAB228p PylRS M. ...mazei Bacterial Ahmed Badran 209743 pAB228p4 PylRS M. mazei Bacterial Ahmed Badran 209744 pAB228v TrpRS...cerevisiae Bacterial Ahmed Badran 209745 pAB228v6 TrpRS S. cerevisiae Bacterial Ahmed Badran 209746 pAMC070n1a12...acids. Expanding the Genetic Code In protein translation, an aminoacyl-tRNA synthetase (aaRS) loads its...Barrick 122650 Mm-PylRS-AF/Pyl-tRNACUA PylRS M. mazei trans-cyclooct- 2-ene-lysine (TCOK) Mammalian TAG Howard...

...from Addgene's viral service encoding tet-off transactivators and tools for affinity purification (TRAP)....Packaging Service: Molecular Tools Tetracycline Transactivators Affinity Purification Neurophysiology Cell ...Cell Ablation Tetracycline Transactivators and Inducible Tools These AAV encode tetracycline-inducible tools...tools/controls and tetracycline transactivators that can be used with tetracycline (tet)-inducible expression...CAG-driven, constitutive Expression of the tet-off transactivator (tTA) 2 Gradinaru 99120 pAAV-ihSyn1-tTA Inducible... promoter (ihSyn) Expression of the tet-off transactivator (tTA) with a positive feedback loop for amplified...ihSyn) Cre-dependent expression of the tet-off transactivator (tTA) with a positive feedback loop for amplified...

...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...(see paper) Transfection none, need Cas9 plasmid Calarco 47943 pMB70 cU6 BsaI Transfection none, need ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined... cut Goldstein 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung 46170 PU6::klp... plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold...sgRNA-scaffold SP6 AflII In vitro transcription none, need Cas9 plasmid Sternberg 177783 L4440_BioBrick-...

...reasons: Transfer plasmids with sequence from the 5′ to the 3′ ITR exceeding 4.7 kb Transfer plasmids ... Terms of Purchase and the applicable material transfer agreements (MTAs) and limited use label licenses...through our Packaged on Request service, Addgene guarantees a titer of at least 4 x 10 12 GC/mL for the AAV1... you know that we don’t expect to produce our guaranteed titer, and will give you the option to proceed...proceed with your order, or not, without a guaranteed titer. Any titer expectations communicated by email ...email will supersede the general guarantees above. We will have our expert viral vector production team carefully...our viral production does not yield the titer guaranteed here or by email, Addgene will contact you to...

... bind transcriptional co-activators and modulate transcription. The list of p53 transcriptional targets...collection of plasmids for the p53 pathway. p53 is a transcription factor and tumor suppressor activated by cellular...p53 Gene List References Background p53 is a transcription factor and tumor suppressor activated by cellular...mutations interfere with p53’s ability to activate transcription, and they also have a dominant negative effect... receptor superfamily, member 10b E2F-1 E2F transcription factor 1 Fas Fas cell surface death receptor... Tyner SD, Venkatachalam S, Choi J, Jones S, Ghebranious N, Igelmann H, Lu X, Soron G, Cooper B, Brayton... p53 mutations in human cancers. Hollstein M, Sidransky D, Vogelstein B, Harris CC. Science. 1991 Jul ...

...efficiently block transcription (in bacteria, ~300-fold repression) by interfering with transcriptional elongation...for selection or fluorescent gating of transiently transfected cells. Co-expression of both plasmids in...elongation, RNA polymerase binding, or transcription factor binding. The binding specificity is determined jointly...the nuclease-deficient mutant could silence transcription on the gene expression level without genetically...CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson MH, Morsut..., Liu Z, Brar GA, Torres SE, Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, Weissman JS...

...Link opens in a new window) . Transfections are performed using the transfer plasmid, a plasmid encoding...cell cultures are inspected for sterility. Transducibility Some viral vectors are tested in vitro and ...section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634...filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then aliquoted, frozen, ...opens in a new window) ). Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector,... taken off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching...integrated provirus using PCR. This assay involves transducing cells with serial dilutions of the lentiviral...

...CRISPR knockout library targeting human transmembrane transporters. Feedback Please send your feedback to...carrier proteins (SLCs) are the largest family of transporters in the human genome with approximately 450 identified... Medicines Initiative 2 Joint Undertaking under grant agreement No 777372. This Joint Undertaking receives...allow a fast and efficient sub-cloning into a wide range of Gateway-compatible pDEST vectors. ID Plasmid ...overexpression library of human SLC proteins. Human Transporter Knockout Libraries 213695 221409 221410 Superti-Furga...

... Plasmids expressing Cas12a/Cas13a, Reverse Transcriptases, etc. that may be useful for detection assays...site for furin proteases.( 3 ) BSG - (CD147), transmembrane glycoprotein of the immunoglobulin superfamily...proteolysis of SARS-CoV Spike protein, activating its membrane fusion function.( 5 ) TACE - (ADAM17) a metalloprotease... method termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR). Mammoth Biosciences has published...2020.06.23.166397 (Link opens in a new window) reverse transcriptase, protease, and Bst polymerase constructs now...unpublished) Taq polymerase and MMLV reverse transcriptase expression plasmids from the Drew Endy and Philippa...2020.03.29.013342 (Link opens in a new window) Reverse Transcriptase plasmids from Andrew Ellington's lab. High-surety...

...identified by depositors and/or when BRAIN Initiative grants are noted in publications associated with Addgene...-GfaABC1D-mCherry-hPMCA2w/b Expresses a plasma membrane calcium pump hPMCA2w/b Baljit Khakh 112000-AAV1...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135421-AAV1 pAAV-syn-FLEX-axon-jYCaMP1s...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135424-AAV1 pAAV-syn-jYCaMP1s...Synapsin1 promoter, for AAV production or direct transfection, slow variant Kaspar Podgorski 135630-AAV1 pAAV-S5E2...interneurons. For use in combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent... expressed under the mammalian promoter (EF1a) Francois St-Pierre 179460-AAV1 pAAV-EF1a-DIO-JEDI-2P-WPRE...

...extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled with the antibody and a species-specific...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light...anti-GFP antibody), the target antigen is first transiently expressed in mammalian cells. To learn which ... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified...

...with a recognizable sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to... is based on the human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein, and is suitable...Vector SNAP-tag Empty backbone for stable and transient mammalian expression 101133 pSNAP-CaaX Control...Control Plasmid SNAP-tag Control for endomembrane (CaaX box) localization 101129 pSNAPf-Cox8A Control Plasmid...Vector CLIP-tag Empty backbone for stable and transient mammalian expression 101134 pCLIPf-H2B Control...pMCP-tag (m) Vector ACP/MCP-tag Empty backbone for transient mammalian expression 101126 pACP-tag (m)- 2 Vector...Vector ACP/MCP-tag Empty backbone for stable and transient mammalian expression 101132 pMCP-GPI Control Plasmid...

...incorporated into the DNA by reverse transcription. The edited DNA strand displaces the unedited 5’ flap and...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...replace” gene editing method in which a reverse transcriptase (RT) is fused to the C terminus of Cas9 H840A...the target sequence where it nicks the non-target strand and generates a 3’ flap. The 3’ flap binds to the...Moloney murine leukemia virus (M-MLV) reverse transcriptase. Engineering of the PE enzyme and related components...including exploring new sources for the reverse transcriptase. Many PE enzymes and related tools are available...

...tools that effectively transduce many mammalian cell types. AAV is used in a wide range of experiments, both...of the applicable transfer plasmid. We will help you gain legal usage of the transfer plasmid from the ...service lets scientists choose from over 3,000 transfer plasmids to be packaged in their choice of five...chemogenetic, optogenetic, recombinase, and biosensor transfer plasmids in our repository are eligible for this...several important steps: MTA facilitation for the transfer plasmid, including legal approval and an implementation...

...including dual-transgenic recombinase-expressing mouse lines 2–9 and combinations of transgenic recombinase-expressing... Cre AND Flp, here expressed through a double transgenic animal strategy (PV-2a-Cre; SOM-IRES-Flp). Image...in a new window) of Flp-recombinase-expressing transgenic mouse and rat lines. The Fenno Lab maintains ... Rayport S. 2018. Dopamine neuron glutamate cotransmission evokes a delayed excitation in lateral dorsal...Neurons in Barrel Cortex Reveals Local and Long-Range Circuit Motifs. Cell Rep. 28(13):3450-3461. PubMed...Krook-Magnuson E. 2019. A novel population of long-range inhibitory neurons. Preprint (Link opens in a new...PubMed (Link opens in a new window) Mandelbaum G, Taranda J, Haynes TM, Hochbaum DR, Huang KW, Hyun M, Umadevi...

....AX with GRE or transgenic animal Oligodendrocytes - AAV-PHP.eB with GRE or transgenic animal Brain vascular....AX with GRE or transgenic animal L5 & Inhibitory - AAV-PHP.eB with GRE or transgenic animal L2/3 & L4...AAV.CAP-B10 with GRE or transgenic animal Excitatory - AAV.PHP.N with GRE or transgenic animal For PNS applications... pAAV-S5E2-GFP-fGFP E2 regulatory element GFP, membrane-targeted GFP Control Dimidschstein 192552 pAAV-CAG... viral prep format. Please note this does not guarantee viral vector packaging service, but lets us know...