LRG1 CRISPR Activation Plasmid (h): sc-405371-ACT

Leucine-rich alpha-2-glycoprotein 1 (LRG1) is a secreted, leucine-rich repeat protein that modulates extracellular signaling in vascular and inflammatory microenvironments. In human cells, LRG1 influences endothelial behavior and tissue remodeling, with well-described links to TGF-β pathway context and alterations in angiogenic signaling outputs. Its expression is regulated during immune activation and stress responses, and elevated LRG1 has been reported across multiple pathological settings involving aberrant neovascularization and chronic inflammation. These properties make LRG1 a useful target for dissecting crosstalk between cytokine signaling, extracellular matrix dynamics, and vascular cell state transitions.

LRG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LRG1 expression without altering the underlying DNA sequence.

LRG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LRG1 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 LRG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LRG1 locus and enabling the study of LRG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRG1 pathway restoration in tumor cells with silenced or reduced LRG1 expression.

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