GP-39 CRISPR Activation Plasmid (h): sc-402147-ACT

CHI3L1 encodes glycoprotein GP-39 (YKL-40), a secreted chitinase-like protein that lacks enzymatic chitinase activity but modulates extracellular matrix remodeling, cell adhesion, and inflammatory signaling. GP-39 is produced by macrophages, neutrophils, fibroblasts, and multiple stromal and epithelial compartments, and it influences chemotaxis, angiogenic programs, and tissue repair responses. It is commonly studied in pathways linked to innate immune activation, cytokine networks, and fibrotic remodeling, including MAPK/ERK, PI3K/AKT, and NF-κB-associated transcriptional responses. Altered CHI3L1/GP-39 expression is associated with chronic inflammation, fibrosis, and tumor-associated microenvironment biology, making it relevant for mechanistic studies of immune–stromal crosstalk and extracellular matrix dynamics.

GP-39 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHI3L1 expression without altering the underlying DNA sequence.

GP-39 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHI3L1 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 CHI3L1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GP-39 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHI3L1 locus and enabling the study of GP-39-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GP-39 pathway restoration in tumor cells with silenced or reduced CHI3L1 expression.

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