PIWIL2 CRISPR Activation Plasmid (h): sc-402211-ACT

PIWIL2 (HIWI2) encodes a PIWI subfamily Argonaute protein that binds piRNAs and contributes to small RNA–guided silencing of transposable elements, supporting genome integrity in germline and germline-like contexts. Through participation in piRNA biogenesis and epigenetic regulation, PIWIL2 influences chromatin state, DNA methylation patterns, and post-transcriptional control of target RNAs. Aberrant PIWIL2 expression has been reported in multiple cancer types and is frequently studied in connection with tumor-associated stem-like phenotypes, proliferation programs, and resistance-associated signaling networks. These features make PIWIL2 a useful node for dissecting RNA-mediated gene regulation and genome defense pathways in human cells.

PIWIL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIWIL2 expression without altering the underlying DNA sequence.

PIWIL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIWIL2 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 PIWIL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PIWIL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIWIL2 locus and enabling the study of PIWIL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PIWIL2 pathway restoration in tumor cells with silenced or reduced PIWIL2 expression.

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