LRP5 CRISPR Activation Plasmid (h): sc-401331-ACT

LRP5 (low-density lipoprotein receptor–related protein 5) is a single-pass transmembrane co-receptor that regulates canonical Wnt/β-catenin signaling by partnering with Frizzled receptors to control ligand-dependent signal transduction. Through modulation of β-catenin stabilization and TCF/LEF-dependent transcription, LRP5 influences osteoblast differentiation, bone mass accrual, and broader programs governing cell fate, proliferation, and tissue homeostasis. LRP5 also participates in crosstalk with extracellular antagonists such as DKK and sclerostin that tune Wnt pathway sensitivity at the cell surface. Genetic and functional perturbation of LRP5 is linked to disorders of bone density and ocular vascular development, making it a key node for studying Wnt-driven phenotypes in human cells.

LRP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LRP5 expression without altering the underlying DNA sequence.

LRP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LRP5 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 LRP5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LRP5 locus and enabling the study of LRP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRP5 pathway restoration in tumor cells with silenced or reduced LRP5 expression.

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