TEF-4 CRISPR Activation Plasmid (h): sc-402319-ACT

Human TEAD2 encodes the TEF-4 transcription factor, a TEA/ATTS domain DNA-binding protein that partners with YAP/TAZ to control gene programs governing proliferation, survival, and lineage specification. TEF-4 integrates signals from the Hippo pathway and coordinates enhancer and promoter activity to regulate cell-cycle progression, cytoskeletal dynamics, and epithelial–mesenchymal plasticity. Dysregulated TEAD2/TEF-4 activity has been linked to aberrant growth control and altered differentiation states observed across multiple tumor contexts and developmental disorders, making it a useful node for dissecting transcriptional circuit dependencies. As a pathway effector, TEF-4 provides a tractable handle to study how upstream mechanical and kinase cues are converted into cell-state transitions.

TEF-4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TEAD2 expression without altering the underlying DNA sequence.

TEF-4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TEAD2 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 TEAD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TEF-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TEAD2 locus and enabling the study of TEF-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TEF-4 pathway restoration in tumor cells with silenced or reduced TEAD2 expression.

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