PRAME CRISPR Activation Plasmid (h): sc-404682-ACT

PRAME (preferentially expressed antigen in melanoma) encodes a cancer-testis antigen that functions primarily as a transcriptional regulator, best known for repressing retinoic acid receptor signaling and attenuating retinoid-driven differentiation programs. By modulating nuclear receptor–dependent transcription and associated epigenetic control, PRAME can influence cell cycle progression, lineage commitment, and apoptosis-related gene expression. Aberrant PRAME expression is reported across multiple tumor contexts and is frequently used as a molecular marker in studies of oncogenic state, immune recognition, and tumor cell plasticity. In human cell models, PRAME perturbation supports investigation of how altered transcriptional networks and differentiation cues contribute to malignant phenotypes and antigen expression profiles.

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

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

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