...so cells must be lysed or the substrate must be delivered intracellularly in order to measure luminescence...Bioluminescence Resonance Energy Transfer (BRET) CalFlux VNT : An intracellular calcium sensor created by fusing...including transcriptional reporters for NF-kb, TGF-b, c-Myc, p53 and MAPK/JNK transcriptional reporters... of luciferase plasmids used for assays of transcriptional regulation and biosensors. Plasmid...insects. Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust... protein half-lives, making them useful as transcriptional reporters. However, they are not secreted, ...regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these...

...easily and inexpensively generate R-mAbs by transiently transfecting the plasmids into mammalian tissue culture... Anti-Pan-Neurofascin (extracellular) [A12/18R] Pan-Neurofascin (extracellular) Rat Mouse IgG2a 177439...Anti-Kv1.4 K+ channel (extracellular) [L71/5R] Kv1.4 K+ channel (extracellular) Rat Mouse IgG2a 177489...Mouse IgG2a 177564 Anti-ADAM22 (extracellular) [N57/2R] ADAM22 (extracellular) Mouse Mouse IgG2a 177565 Anti-GluN2B...channel Rat Mouse IgG2a 188161 Anti-TRAAK K+ channel [N445/27R] TRAAK K+ channel Mouse Mouse IgG2a 188162...Anti-Kv1.4 K+ channel (extracellular) [L71/22] Kv1.4 K+ channel (extracellular) Rat Mouse IgG2a 190292...N382/14] MFF Human Mouse IgG2a 190320 Anti-TRAK2 [N390/43] TRAK2 Human Mouse IgG2a 190321 Anti-Navbeta2 Na...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ... dU6 BspQI Transfection Puromycin yes, cut Ji-Long Liu U6-BbsI-crRNA dU6 BbsI 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...non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants...

...Plant transcription factors for Agrobacterium-mediated transient expression or plant transformation - Silin...parts for assembling plant transformation vectors - Daniel Voytas. Transcription Factors The following collections...expression in plants: Maize Transcription Factor ORFeome - John Gray. Rice Transcription Factor ORFeome - John...Plant Base Editors Save Time with Transient Plant Leaf Transformations RNA Interference in Plant Biology...Figshare - Plant Illustrations (Link opens in a new window) - Collection of illustrations and pictures of... Addgene Blog: Plant Biology CRISPR Cloning Transcription Factors RNAi Empty Backbones Fluorescent Proteins...Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes...

...protein translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery... Mehl 85496 pDule-Tet2.0 Tetrazine2.0 tRNA synthetase M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan Mehl...Mehl 85497 pDule2-Tet2.0 Tetrazine2.0 tRNA synthetas M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan Mehl...barkeri Tetrazine 3.0 Bacterial TAG Ryan Mehl 174081 pAcBac1-NES-Flag-R2-84-MbRS Tetrazine3.0 NES-Flag-R2...ATG David Tirrell Browse Strains The table below highlights bacteria strains that have been modified to...Addgene's guide to plasmids and strains useful for expanding the genetic code in bacteria and mammalian...Rinehart Lab reagents tRNA/Synthetase Pairs Bacterial Strains Codon Modified Constructs The genetic code for ...

... recoded strain of E. coli ( 68306 ) with the optimized phosphoserine orthogonal translation system ( ...Plasmid 69118 E17TAG GFP zeo resistance Strain 34929 BL21ΔserB Strain 52055 EcAR7 Plasmid 112031 pNAS1b split...phosphorylation is one of the most abundant forms of posttranslational modifications in cells and research into its..., Science 2013) and improving the orthogonal translation system to make singly or multiply phosphorylated...pSerOTS. Recently the Rinehart lab published a new strain of recoded E.coli (rEcoli XpS 192872 ) and an optimized... optimized phosphoserine orthogonal translation system (pSerOTS-C1* (V70) 188537 ). These reagents are...phosphorylation sites (“phosphosites”) consist of a central phosphoserine site flanked by 15 amino acids on...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. Plasmid Promoter PI Publication Nick...

...inversion, deletion and translocation. Inversion: If the loxP sites are on the same DNA strand and are in opposite... maintained). Translocation: If the sites are on separate DNA molecules, a translocation event is generated...85577 pTC-CMV-Tet CreER expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian... pTC-ApoE-Tet CreER expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian...plasmids can be useful for lineage tracing, evaluating infection/transfection efficiency, selecting desired... Mammalian Luo 47117 pT3TS-Cre for in vitro transcription of Cre lac Bacterial Ekker 47551 pDD104 (Peft...promoter none AAV Simpson 49455 pSH62-EBD Cre-EBD - Estradiol-dependent control Gal1 Yeast Gartenberg 50797 ...

...Tools Tetracycline Transactivators Affinity Purification Cell Ablation Tetracycline Transactivators and ...These AAV encode tetracycline-inducible tools/controls and tetracycline transactivators that can be used...service encoding tet-off transactivators and tools for affinity purification (TRAP). Viral...used with tetracycline (tet)-inducible expression systems . ID Name Expression System Activity Serotype... 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...Gradinaru 117383 TRE-DIO-eYFP Cre-dependent and Tetracycline-inducible Cre-dependent, Tet-inducible expression...

...- Tatjana Sauka-Spengler Lab Lineage Tracing Cell lineage tracing in multicellular systems typically relies...Lewis Lab PhOTO Zebrafish: A Transgenic Resource for In Vivo Lineage Tracing during Development and Regeneration...Periklis Pantazis Lab Multibow: digital spectral barcodes for cell tracing - Sean Megason Lab Zebrabow - Albert... (~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...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. CRISPR-X, a base-editing system from...

...diluting or concentrating to a final concentration of 0.9–1.1 mg/mL. Antibody concentration is confirmed...Blotting (WB) Proteins are extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled...of co-purified plasmid from transfection. This residual plasmid is extracted and amplified with primers...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light... the buffer can be removed by dialysis or gel filtration. Titer and Purity Antibodies are initially titered...serial dilutions of a standard antibody of known concentration and stained with Coomassie blue. Protein band...standard curve is generated. The final antibody concentration is calculated from the standard curve. Using...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...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...base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. Plasmid Gene/Insert Promoter Selectable...

...reverse transcriptase and integrase. Once in a target cell, the virus uses the reverse transcriptase in tow...long-term transgene expression. Some disadvantages of γ-retroviruses are that they can only transduce dividing...Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral...Plasmid for transgene expression; also expresses luciferase Lowe 9044 pMIG MSCV Plasmid for transgene expression...Addgene, including popular packaging, envelope, and transfer plasmids. Viral...of interest? Search our database for retroviral transfer plasmids that have inserts. ID Plasmid 5'LTR Description... Description PI 1764 pBABE-puro MoMLV For transgene expression, with Puromycin selection (for additional...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...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...base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. Plasmid Gene/Insert Promoter Selectable...

...Link opens in a new window) . Transfections are performed using the transfer plasmid, a plasmid encoding...are purified by iodixanol gradient ultracentrifugation and concentrated to purity and titers adequate for...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...cleared by low-speed centrifugation and then by filtration through a 0.45 µm polyethersulfone (PES) membrane... been stored at -80 °C and thawed. Lentivirus titration is performed using a 293T cell line in the presence...using a lentiviral droplet digital PCR (ddPCR) titration protocol (modified from Wang et al. (2018) (Link...

...information on substrates and synthases needed for use of these tags. Plasmids Addgene ID Plasmid Substrate Feature...simultaneously with dual labeling when used with two substrates Can be used in live or fixed cells, and for internal... proteins and use enzymatic binding to attach substrate Control plasmids with a recognizable sub-cellular... is based on the human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein, and is suitable...protein tags based on the acyl carrier protein. Substrates are derivatives of Coenzyme A (CoA). These tags...Vector SNAP-tag Empty backbone for stable and transient mammalian expression 101133 pSNAP-CaaX Control...Vector CLIP-tag Empty backbone for stable and transient mammalian expression 101134 pCLIPf-H2B Control...

...interfering RNA) via transfection, or shRNA (short hairpin RNA) via transfection or viral delivery of ...silencing is achieved through the delivery of a double-stranded RNA (dsRNA) that matches the mRNA target sequence...or off PI 11643 pLVCT-tTR-KRAB 2nd generation; Transgene (CAG promoter) ‐ OR ‐ shRNA (H1 promoter when ...Trono 11779 pLVCT-rtTR-KRAB-2SM2 2nd generation; Transgene (CAG promoter) ‐ OR ‐ shRNA (H1 promoter when ...Didier Trono 11644 pLVET-tTR-KRAB 2nd generation; Transgene (hEF-1alpha promoter) ‐ OR ‐ shRNA (H1 promoter...Didier Trono 11642 pLVPT-tTR-KRAB 2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when...Trono 11652 pLVPT-rtTR-KRAB-2SM2 2nd generation; Transgene (hPGK promoter) ‐ OR ‐ shRNA (H1 promoter when...

...Adenoviruses have high transduction efficiency and direct high but transient transgene expression. γ-Retrovirus...virus for gene transfer in clinical trials. These viruses have a relatively high transduction efficiency,...Dividing/Non-dividing Yes Low AAV Transient or Stable* single stranded linear DNA ∼4.5 Kb 18-26 Dividing.../Non-dividing No* Very Low Adenovirus Transient double stranded linear DNA >8 Kb 105 Dividing/Non-dividing...are versatile tools because of their ability to transduce non-dividing cells. These viruses enable long-...immunogencity. These viruses can direct long-term transgene expression, but generally are not permanently ...permanently into the host genome, enabling long term transgene expression. Overview of Viral Plasmids While plasmids...

... of the CRISPR RNA (crRNA) and the trans-activating crRNA (tracrRNA; Hsu et al ., Nature Biotechnology...plasmids #61422 - #61424 were designed for transient transfection : 61422 : Expresses dCAS9-VP64 activator... cassettes to express the CRISPR RNA array and tracrRNA separately (Cong et al ., Science 2013). Additionally...or SpCas9n, D10A nickase) + CRISPR RNA array + tracrRNA: This plasmid contains three expression cassettes...human codon-optimized SpCas9; sgRNA can be co-transfected as PCR amplicons (Ran et al., Nature Protocols...references are below. Return to top Cas9 SAM transcriptional activation plasmids and screening library CRISPR...) is an engineered protein complex for the transcriptional activation of endogenous genes. It consists...