...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...

...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...

...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...

...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. Plasmids In addition to ... 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...

... (~3 months), ease of care and breeding, and transparency of the embryo and larvae, it is easy to culture...standard toolbox of genetic methods, such as Tol2 transposons, Cre-mediated site-specific recombination, morpholinos...emerged as a model system for human disease and translational research, including the study of cancer, inflammation...-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference (CRISPRi...zebrafish - Julian Lewis Lab PhOTO Zebrafish: A Transgenic Resource for In Vivo Lineage Tracing during Development...examples of popular tools to use in zebrafish, ranging from optogenetics to tools for studying diseases...vectors designed to streamline the production of transgenic zebrafish for studying neurodegenerative disorders...

...generate double strand breaks (DSBs). Nick Mutated Cas nickase enzymes generate single-strand DNA breaks. ...deletions. Insert NEW CRISPR transposases insert large fragments of DNA. Transcriptional Regulation Activate dCas9...Prime Edit Cas nickase enzymes fused to reverse transcriptase install new genetic information into a specific..., or fused to a repressor peptide, inhibits transcription. Epigenetics dCas9 fused to an epigenetic modifier...

...CRISPR Blog Posts CRISPR-on is an RNA-guided transcriptional activator system with two components, the dCas9...sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component provides the ...dCas9 activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA ...Gateway donor vectors [48218~48221] and can be transferred to Gateway destination vectors (e.g., pmax-DEST...endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cheng AW, Wang H, Yang H,...H, Shi L, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Cell Res. 2013 Oct;...

...reactivation, RNA editing, gene replacement therapy, RNA trans-splicing, and protein replacement to reverse the...cases Rett syndrome can be inherited. In females, random X inactivation results in approximately half of...disease severity. The MECP2 protein is a global transcriptional regulator of thousands of genes and studies...studies have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...DNA and recruit known co-repressors or other transcriptional modifiers suggests the major function of MECP2...NTD) the M ethyl B inding D omain (MBD) a T ranscriptional R epressor D omain (TRD) the TRD contains a...Developing Human Brain - includes developmental transcriptome information, prenatal LMD microarray data, ISH...