Protamine 2 CRISPR Activation Plasmid (h): sc-402337-ACT

PRM2 encodes protamine 2, a sperm-specific, arginine-rich DNA-binding protein that replaces histones during late spermiogenesis to drive chromatin condensation and nuclear compaction. This transition is a key component of spermatid differentiation and genome packaging, influencing sperm head morphology, DNA integrity, and epigenetic organization in the male germline. Altered PRM2 expression or protamine imbalance is associated with defective chromatin remodeling, increased DNA fragmentation, and impaired semen parameters, making it relevant to studies of male reproductive biology and infertility-associated mechanisms. PRM2 is commonly evaluated alongside other chromatin remodeling factors to probe the molecular control of spermiogenesis and germ cell maturation.

Protamine 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRM2 expression without altering the underlying DNA sequence.

Protamine 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRM2 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 PRM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Protamine 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRM2 locus and enabling the study of Protamine 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Protamine 2 pathway restoration in tumor cells with silenced or reduced PRM2 expression.

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