Laminin β-2 CRISPR Activation Plasmid (h): sc-402612-ACT

LAMB2 encodes laminin β2, a core subunit of laminin heterotrimers within basement membranes that supports tissue architecture and cell–matrix adhesion. Laminin β2 contributes to extracellular matrix assembly and signaling through integrin- and dystroglycan-associated pathways, influencing cytoskeletal organization, cell polarity, and differentiation. In the kidney glomerular basement membrane and at neuromuscular synapses, laminin β2 is important for filtration barrier integrity and synaptic maturation, respectively. Dysregulated or deficient LAMB2 function is linked to basement membrane disorders and is commonly studied in contexts such as podocyte biology, glomerular injury mechanisms, and extracellular matrix remodeling.

Laminin β-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAMB2 expression without altering the underlying DNA sequence.

Laminin β-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAMB2 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 LAMB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Laminin β-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAMB2 locus and enabling the study of Laminin β-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Laminin β-2 pathway restoration in tumor cells with silenced or reduced LAMB2 expression.

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