TM4SF1 CRISPR Activation Plasmid (h): sc-404779-ACT

TM4SF1 (transmembrane 4 L six family member 1) is a tetraspanin-like cell surface glycoprotein enriched in endothelial and epithelial contexts and implicated in regulating cell adhesion, motility, and cytoskeletal remodeling. It organizes membrane microdomains and modulates signaling output from integrins and receptor tyrosine kinases, influencing pathways linked to migration, extracellular matrix interaction, and angiogenic programs. TM4SF1 expression has been associated with invasive phenotypes and vascular remodeling across multiple disease settings, making it a useful node for studying microenvironment-driven changes in cell behavior. As a membrane scaffold protein, TM4SF1 is frequently examined for its contribution to cell–cell and cell–matrix communication and downstream transcriptional responses.

TM4SF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TM4SF1 expression without altering the underlying DNA sequence.

TM4SF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TM4SF1 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 TM4SF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TM4SF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TM4SF1 locus and enabling the study of TM4SF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TM4SF1 pathway restoration in tumor cells with silenced or reduced TM4SF1 expression.

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