CLEAR Reporter Lentivirus

Background

The CLEAR (Coordinated Lysosomal Enhancement and Regulation) gene network is a transcriptional program that governs lysosomal biogenesis, autophagy, and vesicular trafficking. Its master regulator is TFEB (transcription factor EB), a basic helix-loop-helix leucine-zipper transcription factor that binds CLEAR elements in the promoters of genes encoding lysosomal enzymes, lysosomal structural proteins, and core autophagy machinery. TFEB activity is controlled by the mTORC1-AKT axis: under nutrient-rich conditions mTORC1 phosphorylates TFEB and retains it in the cytoplasm, whereas starvation, lysosomal stress, or mTOR inhibition trigger TFEB dephosphorylation and nuclear translocation. Dysregulated TFEB signaling is implicated in lysosomal storage disorders, neurodegeneration, and cancer.

Product Description & Applications

The CLEAR Reporter Lentivirus is a lentiviral transcription-factor reporter system that gives a sensitive fluorescent (GFP or RFP) or luminescent (firefly luciferase) readout of CLEAR gene network activity, reflecting TFEB transcriptional output. Tandem repeats of modified CLEAR elements drive reporter expression upon TFEB activation, enabling detection of autophagy-lysosome pathway induction by stressors, nutrient deprivation, and mTOR-AKT inhibition. The particles are purified by PEG precipitation and sucrose gradient centrifugation, and a constitutively expressed drug-selection marker (puromycin or blasticidin) supports rapid establishment of stable reporter cell lines. The system efficiently transduces difficult-to-transfect cells, including primary and cryopreserved cultures, and supports readout by fluorescence microscopy, flow cytometry, or luminometry for autophagy and protein-homeostasis studies.

About This Product

This reporter lentivirus places a EGFP, Firefly Luc, GFP, Luc, mCherry, Renilla Luc, RFP reporter gene under the control of tandem consensus response elements specific for the CLEAR gene network transcription factor, coupled to a minimal TATA-box promoter and a proprietary upstream enhancer that maximizes signal-to-noise. The constitutively expressed selection marker (Puromycin, Blasticidin) and/or secondary reporter enables stable polyclonal cell line generation and flexible readout by fluorescence microscopy, flow cytometry, or luminometry.

Stable integration via the lentiviral backbone ensures consistent, clonally representative reporter expression in dividing and post-mitotic target cells — including primary T cells, macrophages, organoids, and cryopreserved material — eliminating the variability inherent to transient transfection. The self-inactivating LTR design and third-generation packaging minimize insertional mutagenesis risk and ensure biosafety classification at BSL-2.