ASGPR1/Asialoglycoprotein Receptor 1/ASGR1 CRISPR Activation Plasmid (h): sc-400429-ACT

ASGR1 encodes asialoglycoprotein receptor 1 (ASGPR1), a hepatocyte-enriched C-type lectin that forms a functional receptor complex at the sinusoidal plasma membrane to recognize terminal galactose or N-acetylgalactosamine on desialylated glycoproteins. This receptor mediates clathrin-dependent endocytosis and lysosomal trafficking, supporting serum glycoprotein homeostasis and hepatic clearance of circulating glycoproteins and glycan-modified particles. ASGPR1 activity links glycan recognition to receptor recycling and intracellular sorting pathways, integrating with endocytic machinery and hepatic metabolic processes. Altered ASGR1 expression or function has been associated with changes in lipoprotein handling and cardiometabolic phenotypes, making it relevant for studies of liver-specific uptake, plasma protein turnover, and glyco-immune interactions.

ASGPR1/Asialoglycoprotein Receptor 1/ASGR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ASGR1 expression without altering the underlying DNA sequence.

ASGPR1/Asialoglycoprotein Receptor 1/ASGR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ASGR1 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 ASGR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASGPR1/Asialoglycoprotein Receptor 1/ASGR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ASGR1 locus and enabling the study of ASGPR1/Asialoglycoprotein Receptor 1/ASGR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASGPR1/Asialoglycoprotein Receptor 1/ASGR1 pathway restoration in tumor cells with silenced or reduced ASGR1 expression.

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