TMEFF1 CRISPR Activation Plasmid (h): sc-404818-ACT

Human TMEFF1 (transmembrane protein with EGF-like and two follistatin-like domains 1) encodes a single-pass membrane protein implicated in extracellular ligand interactions and modulation of growth factor signaling. Reported functions link TMEFF1 to regulation of cell proliferation and differentiation programs, with context-dependent effects on receptor tyrosine kinase pathways such as EGFR/ERK and PI3K–AKT signaling. TMEFF1 expression is frequently dysregulated across multiple tumor types and has been associated with altered epithelial cell states, invasiveness, and microenvironmental signaling. These attributes make TMEFF1 a useful target for studying signal integration at the plasma membrane and transcriptional consequences of growth factor pathway perturbation.

TMEFF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TMEFF1 expression without altering the underlying DNA sequence.

TMEFF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TMEFF1 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 TMEFF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TMEFF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TMEFF1 locus and enabling the study of TMEFF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TMEFF1 pathway restoration in tumor cells with silenced or reduced TMEFF1 expression.

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