Laminin α-5 CRISPR Activation Plasmid (h): sc-401168-ACT

LAMA5 encodes laminin α-5, a core subunit of laminin-511 and laminin-521 heterotrimers that organize basement membranes and regulate epithelial and endothelial adhesion, polarity, and differentiation. Through interactions with integrins, dystroglycan, and other extracellular matrix receptors, laminin α-5 contributes to focal adhesion signaling and downstream pathways such as FAK/Src, PI3K–AKT, and MAPK that coordinate migration and survival. LAMA5-rich matrices shape tissue architecture in skin, kidney glomerulus, lung, and vasculature, influencing barrier function and morphogenesis. Dysregulated laminin α-5 deposition or signaling is associated with altered basement membrane integrity and has been studied in contexts including fibrosis, inflammatory remodeling, and tumor invasion microenvironments.

Laminin α-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMA5 expression without altering the underlying DNA sequence.

Laminin α-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMA5 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 LAMA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Laminin α-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMA5 locus and enabling the study of Laminin α-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Laminin α-5 pathway restoration in tumor cells with silenced or reduced LAMA5 expression.

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