LGR5 CRISPR Activation Plasmid (h): sc-400726-ACT

LGR5 (leucine-rich repeat–containing G protein-coupled receptor 5) is a seven-transmembrane receptor that functions as a stem and progenitor cell marker in multiple epithelia, including intestinal crypts and hair follicles. It serves as a high-affinity receptor for R-spondins and potentiates canonical WNT/β-catenin signaling by stabilizing Frizzled/LRP receptor complexes, thereby supporting self-renewal, proliferation, and tissue regeneration programs. Through modulation of WNT-driven transcriptional networks, LGR5 helps regulate differentiation trajectories and niche interactions in developing and adult tissues. Dysregulated LGR5 expression and WNT pathway hyperactivation are frequently associated with tumor biology and are used to interrogate cancer stem-like states and lineage plasticity in human model systems.

LGR5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LGR5 expression without altering the underlying DNA sequence.

LGR5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LGR5 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 LGR5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LGR5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LGR5 locus and enabling the study of LGR5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LGR5 pathway restoration in tumor cells with silenced or reduced LGR5 expression.

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