Peg10 CRISPR Activation Plasmid (m): sc-431278-ACT

Mouse Peg10 (paternally expressed gene 10) encodes a domesticated retrotransposon-derived protein with Gag- and Pol-like features that supports cell survival, proliferation, and lineage-specific differentiation. Peg10 is regulated by genomic imprinting and contributes to developmental programs, including placental and embryonic growth, through transcriptional and post-transcriptional control of gene expression. At the cellular level, Peg10 has been linked to pathways governing apoptosis resistance and proteostasis, and its dysregulation is frequently studied in the context of oncogenic transformation, tissue remodeling, and aberrant growth signaling. These properties make Peg10 a relevant target for investigating imprinted gene networks, developmental biology, and disease-associated changes in gene regulatory circuitry.

Peg10 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Peg10 expression without altering the underlying DNA sequence.

Peg10 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Peg10 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 Peg10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Peg10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Peg10 locus and enabling the study of Peg10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Peg10 pathway restoration in tumor cells with silenced or reduced Peg10 expression.

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