CD24 CRISPR Activation Plasmid (h): sc-418762-ACT

CD24 encodes a glycosylphosphatidylinositol (GPI)-anchored, heavily glycosylated cell surface protein that modulates cell–cell and cell–matrix interactions and shapes immune signaling. CD24 participates in processes including lymphocyte activation, adhesion dynamics, and regulation of inflammatory responses through interactions with sialic acid–binding immunoglobulin-like lectins, influencing innate immune checkpoints and downstream signaling programs. Its expression is developmentally regulated and often linked to epithelial and hematopoietic lineage states, making it a useful marker for cellular differentiation and phenotypic heterogeneity. Altered CD24 abundance has been associated with immune dysregulation and tumor biology contexts, supporting mechanistic studies of immune evasion, metastasis-associated traits, and microenvironmental crosstalk in model systems.

CD24 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD24 expression without altering the underlying DNA sequence.

CD24 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD24 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 CD24 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD24 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD24 locus and enabling the study of CD24-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD24 pathway restoration in tumor cells with silenced or reduced CD24 expression.

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