TEF-1 CRISPR Activation Plasmid (h): sc-402607-ACT

Human TEAD1 encodes the TEF-1 transcription factor, a TEA/ATTS domain DNA-binding protein that partners with YAP/TAZ to regulate gene programs controlling cell proliferation, survival, and lineage specification. TEF-1 functions as a nuclear effector of the Hippo signaling pathway and integrates inputs from mechanotransduction and growth factor signaling to modulate enhancer and promoter activity. Dysregulated TEAD–YAP/TAZ transcriptional output has been associated with oncogenic transcriptional states, fibrosis-associated remodeling, and altered developmental patterning, making TEAD1 a key node in studies of tissue growth control. TEAD1 also contributes to muscle and cardiovascular gene expression networks, supporting investigations into differentiation and stress-response transcriptional circuitry.

TEF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TEAD1 expression without altering the underlying DNA sequence.

TEF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TEAD1 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 TEAD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TEF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TEAD1 locus and enabling the study of TEF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TEF-1 pathway restoration in tumor cells with silenced or reduced TEAD1 expression.

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