CD42b CRISPR Activation Plasmid (h): sc-401273-ACT

GP1BA encodes the platelet surface glycoprotein Ib alpha chain (CD42b), an essential component of the GPIb-IX-V receptor complex that mediates platelet tethering to von Willebrand factor under high shear conditions. This receptor initiates signaling events that support platelet adhesion, activation, and thrombus formation, interfacing with cytoskeletal remodeling and downstream coagulation-related processes. Altered GP1BA expression or function is linked to inherited platelet adhesion defects such as Bernard–Soulier syndrome and is widely studied in the context of bleeding diatheses and thrombotic risk. Because CD42b is largely restricted to megakaryocytes and platelets, it serves as a useful marker and functional node for investigating platelet biogenesis and hemostatic signaling pathways.

CD42b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GP1BA expression without altering the underlying DNA sequence.

CD42b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GP1BA 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 GP1BA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD42b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GP1BA locus and enabling the study of CD42b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD42b pathway restoration in tumor cells with silenced or reduced GP1BA expression.

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