LRP1B CRISPR Activation Plasmid (h): sc-404088-ACT

LRP1B (LDL receptor related protein 1B) is a large endocytic receptor in the LDL receptor family that participates in receptor-mediated internalization of extracellular ligands and modulation of cell-surface protease and growth factor availability. Through its effects on endocytosis and membrane protein turnover, LRP1B can influence lipid handling, extracellular matrix remodeling, and signaling pathways linked to cell adhesion and migration. Altered LRP1B expression or genomic disruption has been reported across multiple cancer-associated datasets and is studied in relation to tumor evolution, invasion, and immune-related features, with additional interest in neuronal and cardiometabolic biology. As a result, LRP1B is frequently examined as a regulator of cellular uptake processes and microenvironmental signaling relevant to disease mechanisms.

LRP1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LRP1B expression without altering the underlying DNA sequence.

LRP1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LRP1B 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 LRP1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRP1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LRP1B locus and enabling the study of LRP1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRP1B pathway restoration in tumor cells with silenced or reduced LRP1B expression.

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