PRDM14 CRISPR Activation Plasmid (h): sc-404426-ACT

PRDM14 (PR domain zinc finger protein 14) is a sequence-specific transcriptional regulator that helps control pluripotency programs, epigenetic reprogramming, and germ cell–associated gene expression in human cells. It influences chromatin state through modulation of histone marks and DNA methylation dynamics, coordinating transcriptional networks that govern cell fate decisions and differentiation. PRDM14 activity intersects with pathways that maintain stem-like transcriptional states and regulate lineage commitment, making it relevant for studies of developmental biology and cellular reprogramming. Dysregulated PRDM14 expression has been linked to altered transcriptional circuitry and epigenetic imbalance observed across multiple cancer contexts, supporting its investigation as a node in oncogenic stemness-associated programs.

PRDM14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRDM14 expression without altering the underlying DNA sequence.

PRDM14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRDM14 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 PRDM14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRDM14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRDM14 locus and enabling the study of PRDM14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRDM14 pathway restoration in tumor cells with silenced or reduced PRDM14 expression.

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