Key Advantages:

• 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.
• 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.
• Same Cost For Custom Lentivirus – You can easily request any combination of reporter (GFP/RFP/Luc) and selection marker (puromycin/blasticidin) for this product, without additional cost, by contacting us. To view our complete list of vectors, click here.

Product Data:

Figure 1. HEK293FT cells were co-transfected with FXRE-TAG-Puro plus either control plasmid (Vector-RFP) or expression plasmid for human FXR (FXR-RFP). Fluorescence microscopy images were taken 24 hrs later to assess GFP reporter expression in the live cells. Expression vector lentivirus for FXR is available by clicking here.

Details:

Vector Diagram

Recommended Controls

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.

Renilla Luciferase Internal Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven Renilla luciferase (RLuc).

Publications

Loyo-Celis V, Patel D, Sanghvi S, Kaur K, Ponnalagu D, Zheng Y, Bindra S, Bhachu HR, Deschenes I, Gururaja Rao S, Singh H. Biophysical characterization of chloride intracellular channel 6 (CLIC6). J Biol Chem. 2023 Nov;299(11):105349. doi: 10.1016/j.jbc.2023.105349. Epub 2023 Oct 12. PMID: 37838179; PMCID: PMC10641671.

Gao AY, Diaz Espinosa AM, Nguyen BBN, Link PA, Meridew J, Jones DL, Gibbard DF, Tschumperlin DJ, Haak AJ. Dopamine Receptor D1 Is Exempt from Transforming Growth Factor β-Mediated Antifibrotic G Protein-Coupled Receptor Landscape Tampering in Lung Fibroblasts. J Pharmacol Exp Ther. 2023 Sep;386(3):277-287. doi: 10.1124/jpet.122.001442. Epub 2023 Apr 6. PMID: 37024146; PMCID: PMC10449101.