CD72 CRISPR Activation Plasmid (h): sc-404054-ACT

CD72 is a B cell–restricted type II transmembrane glycoprotein that functions as an inhibitory co-receptor to tune B cell receptor (BCR) signaling thresholds and maintain immune homeostasis. Through its cytoplasmic ITIM motif, CD72 recruits phosphatases such as SHP-1 to dampen proximal kinase cascades and modulate downstream pathways including MAPK and NF-κB, influencing B cell activation, proliferation, and tolerance. CD72 also participates in regulatory interactions with ligands such as SEMA4D/CD100, integrating signals that affect antigen-driven responses and germinal center dynamics. Dysregulated CD72 expression or signaling has been linked to altered B cell function and is relevant to studies of autoimmunity, B cell malignancies, and immune checkpoint regulation in the tumor microenvironment.

CD72 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD72 expression without altering the underlying DNA sequence.

CD72 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD72 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 CD72 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD72 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD72 locus and enabling the study of CD72-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD72 pathway restoration in tumor cells with silenced or reduced CD72 expression.

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