Laminin β-1 CRISPR Activation Plasmid (h): sc-401105-ACT

LAMB1 encodes laminin β1, a core subunit of multiple laminin heterotrimers that form a major structural and signaling component of basement membranes. Laminin β1 supports extracellular matrix assembly, cell adhesion, polarity, and migration through interactions with integrins and other matrix receptors, influencing pathways such as focal adhesion signaling and cytoskeletal remodeling. By shaping basement membrane integrity and cell–matrix communication, laminin β1 contributes to tissue architecture in epithelia, vasculature, and nervous system compartments. Dysregulated LAMB1 expression or basement membrane organization is frequently studied in the context of developmental defects, tumor invasion and metastasis biology, and fibrotic or inflammatory microenvironments.

Laminin β-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMB1 expression without altering the underlying DNA sequence.

Laminin β-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMB1 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 LAMB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Laminin β-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMB1 locus and enabling the study of Laminin β-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Laminin β-1 pathway restoration in tumor cells with silenced or reduced LAMB1 expression.

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