PLAC8 CRISPR Activation Plasmid (h): sc-405093-ACT

PLAC8 (placenta associated 8) encodes a small, cysteine-rich protein implicated in the regulation of cellular differentiation, epithelial–mesenchymal plasticity, and innate immune-related signaling. In human cells, PLAC8 has been linked to modulation of proliferative and stress-response programs, including pathways connected to autophagy, redox balance, and cytoskeletal remodeling. Altered PLAC8 expression has been reported across multiple disease contexts, particularly in cancers where it correlates with changes in invasion, metastasis-associated phenotypes, and tumor microenvironment interactions. These attributes make PLAC8 a useful node for mechanistic studies of transcriptional control, cell state transitions, and context-dependent signaling outputs.

PLAC8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLAC8 expression without altering the underlying DNA sequence.

PLAC8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLAC8 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 PLAC8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLAC8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLAC8 locus and enabling the study of PLAC8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLAC8 pathway restoration in tumor cells with silenced or reduced PLAC8 expression.

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