SMP30 CRISPR Activation Plasmid (h): sc-401934-ACT

Human RGN encodes senescence marker protein 30 (SMP30), a multifunctional cytosolic protein implicated in cellular aging, oxidative stress responses, and calcium homeostasis. SMP30 has been linked to regulation of reactive oxygen species handling, hepatocyte metabolism, and maintenance of membrane stability under stress, integrating with pathways that influence cell survival and tissue resilience. Altered SMP30 expression has been associated with age-related functional decline and metabolic dysregulation in liver and other tissues, making RGN a useful node for mechanistic studies of senescence-associated phenotypes. As a conserved stress-response factor, SMP30 is frequently examined in models of inflammation, redox imbalance, and cellular homeostatic failure.

SMP30 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RGN expression without altering the underlying DNA sequence.

SMP30 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RGN 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 RGN transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMP30 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RGN locus and enabling the study of SMP30-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMP30 pathway restoration in tumor cells with silenced or reduced RGN expression.

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