PRMT5 CRISPR Activation Plasmid (h): sc-401115-ACT

PRMT5 (protein arginine methyltransferase 5) is a type II arginine methyltransferase that catalyzes symmetric dimethylation of arginine residues on histones and diverse non-histone substrates, shaping chromatin structure and transcriptional programs. Through interactions with cofactors such as WDR77/MEP50, PRMT5 influences RNA splicing, ribosome biogenesis, DNA damage responses, and cell-cycle control, linking epigenetic regulation to proteostasis and genome maintenance. PRMT5 activity intersects with pathways governing differentiation and stress signaling, and its dysregulation has been associated with altered transcriptional states observed in multiple cancers and in contexts of neurodevelopmental and immune perturbation.

PRMT5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRMT5 expression without altering the underlying DNA sequence.

PRMT5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRMT5 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 PRMT5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRMT5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRMT5 locus and enabling the study of PRMT5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRMT5 pathway restoration in tumor cells with silenced or reduced PRMT5 expression.

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