TEAD Reporter Lentivirus
Fluorescent TEAD Reporter Lentivirus (Hippo Pathway): High-quality, fluorescent lentiviral transcription factor (TF) reporter system that provides sensitive fluorescent readout for human/mouse TEA domain transcription factor (TEAD) activity in transduced cells. The transcription factor reporter lentivirus is purified by PEG precipitation and sucrose gradient centrifugation, and is ideal for studying TEAD/YAP or Hippo pathway activity in difficult-to-transfect cells including primary and/or thawed cells.
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Ultra-high concentration virus can be provided upon request in your choice of medium and volume for in vivo injection. We can provide custom reporter genes (such as SEAP, YFP, secreted Gaussia Luc, Renilla Luc, etc.) and selection markers (hygromycin, bleomycin, etc.) upon request.
- High Sensitivity – LipExoGen TF Reporter lentiviral particles are made using a novel vector platform based on the third generation system. The transcription factor’s response elements are arranged as DNA tandem repeats upstream of the minimal TATA promoter-driven reporter, and downstream of an optimized minimal enhancer (pc) of the human CMV promoter. When the signal pathway/TF is activated, the mini enhancer synergizes with TF binding to the response elements (up to 8 repeats in some products, depending on strength of reporter activation) to amplify expression of the fluorescent (GFP/RFP) or luciferase (Luc) reporter, with minimal enhancement of background. As a result, the reporter system provides a highly sensitive readout for signaling pathway or specific transcription factor activation in human and mouse cells.
- Easily Establish Stable Reporter Cell Lines – The reporter lentiviral particles are ultra-purified and concentrated to high-titer by PEG purification and sucrose gradient centrifugation to allow for efficient transduction of difficult-to-transfect cells, including primary and/or freeze-thawed cells. Stable cell lines are easily generated through puromycin or blasticidin selection.
- Discovery Made Easy – Signal pathway or specific transcription factor activity can be detected by fluorescence, making LipExoGen TF Reporter lentiviral particles more practical than traditional luciferase reporters and/or biochemical assays. Pathway/TF activation can be readout directly by fluorescence microscopy in living cell cultures, thus paving the way for unexpected discoveries.
- Readout On Flow – Fluorescent reporter activation can also be readout by flow cytometry, providing more versatility in data acquisition for labs with different instruments.
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- Best Value – LipExoGen lentiviral particle products are made using optimized lentiviral vectors developed in-house, which allows us to provide the highest quality products while retaining competitive prices. These high-titer lentiviral particles feature a highly sensitive fluorescent reporter system and have been validated to read out the indicated transcription factor activity.
Figure 1 (thumbnail). HEK293FT cells were co-transfected with TEAD-TAG-Puro plus either control plasmid (pcDNA Vector, left) or expression plasmid for human YAP1 (YAP1, right). Fluorescence microscopy images were taken 24-36 hrs later to assess GFP reporter expression in the live cells.
Figure 2. GFP reporter activation in cells co-transfected with TEAD-TAG-Puro and YAP1-P2A-RFP. HEK293FT cells were co-transfected with TEAD-TAG-Puro plus human YAP1-P2A-RFP (bottom) or pcDNA vector (top). After 24-36 hours, GFP reporter activity of TEAD-TAG-Puro was observed in the living cells with overexpression of human YAP1 (RFP).
Figure 3. GFP or RFP reporter co-expression with Firefly Luc HEK293FT cells were co-transfected with plasmid constructs for a TEAD dual reporter plus either vector or YAP1-HA for 36 h. Fluorescence microscopy images were acquired in the live cells to observe fluorescent reporter expression (top panels). The figure on the left shows the data for cells transfected with the construct for TEAD-GFP-Luc-Puro (LTV-0016-9S), while the figure on the right shows the data for TEAD-RFP-Luc-Puro (LTV-0016-12S). To visualize Firefly luciferase expression, the cells were then fixed with methanol and stained with anti-Luc-Alexa-594 or -488 antibodies, followed by fluorescence microscopy. The fluorescent (GFP or RFP) reporter is separated from downstream Luciferase reporter by P2A, but driven by the same YAP1-activated TEAD DNA-binding elements. The co-expression of fluorescent and luminescent reporters shows that the reporter can readout fluorescence and luciferase simultaneously in response to Hippo pathway activation.
Have questions about this product? Send us a form and we’ll reply the same day: Contact Us
|TEAD response element|
|Monitor Hippo activity. The fluorescent reporter enables convenient readout using flow cytometry, fluorescence microscopy, etc.|
Renilla Luciferase Internal Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven Renilla luciferase (RLuc).
Reporter Negative Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven reporter. The construct is the same as the TF Reporters except that it lacks the transcriptional response elements which drive signal pathway/TF-specific reporter expression. The reporter negative control lentiviral particles allow to establish a baseline for background reporter activity and determine specificity of any treatments to activate the reporter.
Reporter Positive Control Lentivirus – Ready-to-transduce lentiviral particles with constitutive expression of the reporter. The reporter positive control lentivirus is useful for transduction optimization studies, especially if the cells are very sensitive or difficult-to-transduce.
If you require a modification to one of our products, such as a change in reporter or other vector component, contact us. We provide fast and efficient services for custom cloning and lentiviral particles on demand, usually for the same or comparable price as the listed item.
Or, send us your cells and we will establish a stable reporter cell line for you using this product. Learn more.
|TEA domain transcription factor 1|
|Homo sapiens/mus musculus|
|Alias||Member 1 (SV40 Transcriptional Enhancer Factor); Transcriptional Enhancer Factor TEF-1; Transcriptional Enhancer Factor 1; Transcription Factor 13; Protein GT-IIC; NTEF-1; TCF-13; TEAD-1; TCF13; Atrophia Areata, Peripapillary Chorioretinal Degeneration; TEA Domain Family Member 1; TEF-1; REF1; TEF1; AA|
|Gene IDs||HGNC:HGNC:11714 Ensembl:ENSG00000187079 MIM:189967|
|Entrez Gene Summary||“This gene encodes a ubiquitous transcriptional enhancer factor that is a member of the TEA/ATTS domain family. This protein directs the transactivation of a wide variety of genes and, in placental cells, also acts as a transcriptional repressor. Mutations in this gene cause Sveinsson’s chorioretinal atrophy. Additional transcript variants have been described but their full-length natures have not been experimentally verified. [provided by RefSeq, May 2010]”|