LSECtin CRISPR Activation Plasmid (h): sc-404538-ACT

Human CLEC4G encodes LSECtin (CLEC4G), a C-type lectin predominantly expressed by liver sinusoidal endothelial cells that functions in glycan-dependent recognition and adhesion. Through its carbohydrate recognition domain, LSECtin participates in endocytosis and modulation of cell–cell interactions at the hepatic sinusoid, shaping antigen handling and immune cell trafficking in the liver microenvironment. CLEC4G/LSECtin activity intersects with lectin–glycoprotein recognition networks and innate immune signaling that influence inflammatory responses and tissue homeostasis. Altered expression has been reported in contexts of hepatic inflammation, fibrosis, and tumor-associated immune modulation, making it relevant for mechanistic studies of liver disease biology.

LSECtin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLEC4G expression without altering the underlying DNA sequence.

LSECtin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLEC4G 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 CLEC4G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LSECtin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLEC4G locus and enabling the study of LSECtin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LSECtin pathway restoration in tumor cells with silenced or reduced CLEC4G expression.

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