TFE3 CRISPR Activation Plasmid (m): sc-437344-ACT

Mouse Tfe3 encodes the transcription factor TFE3, a MiT/TFE family member that binds E-box motifs to coordinate programs controlling lysosomal biogenesis, autophagy, cellular metabolism, and stress adaptation. TFE3 integrates nutrient-sensing signals, including mTORC1-dependent regulation, to modulate transcriptional outputs linked to endolysosomal trafficking and mitochondrial homeostasis. Dysregulated TFE3 activity has been associated with altered catabolic flux, aberrant immune and inflammatory signaling, and oncogenic transcriptional reprogramming in models of tumorigenesis. In biomedical research, Tfe3 is commonly studied for its role in TFEB/TFE3-driven lysosomal gene networks, metabolic remodeling, and context-dependent effects on proliferation and differentiation.

TFE3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tfe3 expression without altering the underlying DNA sequence.

TFE3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tfe3 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the Tfe3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TFE3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tfe3 locus and enabling the study of TFE3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TFE3 pathway restoration in tumor cells with silenced or reduced Tfe3 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.