Glycophorin A CRISPR Activation Plasmid (h): sc-417801-ACT

GYPA encodes glycophorin A, a highly abundant sialoglycoprotein of the human erythrocyte membrane that contributes to red blood cell surface charge, membrane stability, and interactions with the extracellular milieu. As a major carrier of MNS blood group antigens, GYPA participates in defining erythrocyte antigenic identity and influences processes such as cell–cell recognition and immune compatibility. Variation in GYPA expression or sequence is relevant to hematologic phenotypes and transfusion immunology, and it is frequently used as a lineage marker in studies of erythropoiesis and red cell differentiation. Glycophorin A biology is also leveraged to interrogate host–pathogen interactions at the erythrocyte surface and mechanisms governing membrane protein trafficking and glycosylation in erythroid cells.

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

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

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