Laminin γ-1 CRISPR Activation Plasmid (h): sc-401029-ACT

LAMC1 encodes laminin γ1, an essential extracellular matrix glycoprotein chain that assembles with α and β subunits to form multiple laminin isoforms in basement membranes. Laminin γ1 supports cell adhesion, polarity, and migration by engaging integrins and other receptors, coordinating signaling that influences cytoskeletal organization, survival, and differentiation programs. Through its role in basement membrane integrity, LAMC1 contributes to tissue morphogenesis and epithelial–mesenchymal interactions, processes frequently remodeled in fibrosis, tumor invasion, and vascular dysfunction. Dysregulated laminin networks and altered LAMC1 expression have been associated with aberrant extracellular matrix deposition and changes in cell–matrix signaling observed across diverse human diseases.

Laminin γ-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMC1 expression without altering the underlying DNA sequence.

Laminin γ-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMC1 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 LAMC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Laminin γ-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMC1 locus and enabling the study of Laminin γ-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Laminin γ-1 pathway restoration in tumor cells with silenced or reduced LAMC1 expression.

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