PRDM16 CRISPR Activation Plasmid (h): sc-403464-ACT

PRDM16 encodes a PR/SET domain–containing transcriptional regulator that orchestrates cell fate programs by modulating chromatin state and gene expression networks. In human cells, PRDM16 integrates lineage specification with metabolic regulation, influencing pathways linked to mitochondrial biogenesis, oxidative metabolism, and developmental transcriptional circuitry. It participates in transcriptional and epigenetic control processes through interactions with co-regulators that shape enhancer activity and differentiation trajectories. Dysregulated PRDM16 expression or function has been associated with altered adipose biology, cardiometabolic phenotypes, and hematopoietic transcriptional programs, making it a useful node for mechanistic studies of differentiation and metabolism.

PRDM16 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRDM16 expression without altering the underlying DNA sequence.

PRDM16 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRDM16 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 PRDM16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRDM16 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRDM16 locus and enabling the study of PRDM16-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRDM16 pathway restoration in tumor cells with silenced or reduced PRDM16 expression.

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