CD34 CRISPR Activation Plasmid (h): sc-400197-ACT

Human CD34 encodes a heavily glycosylated transmembrane sialomucin that serves as a canonical marker of hematopoietic stem and progenitor cells and is also expressed on vascular endothelial cells. CD34 contributes to cell adhesion and trafficking by modulating interactions with selectins and extracellular matrix components, influencing homing, migration, and niche retention in hematopoietic tissues. Its expression is dynamically regulated during differentiation and is commonly used to define stem-like and progenitor-enriched populations in hematopoiesis and vascular biology. Altered CD34 expression patterns are frequently observed across hematologic and vascular pathologies, supporting its utility for mechanistic studies of lineage state, microenvironmental interactions, and disease-associated remodeling.

CD34 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD34 expression without altering the underlying DNA sequence.

CD34 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD34 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 CD34 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD34 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD34 locus and enabling the study of CD34-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD34 pathway restoration in tumor cells with silenced or reduced CD34 expression.

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