STELLA CRISPR Activation Plasmid (h): sc-403261-ACT

DPPA3 (STELLA) is a germ cell- and early embryo-associated factor implicated in epigenetic regulation during preimplantation development, including maintenance of DNA methylation states and protection of genomic imprints. In human cells, STELLA is linked to chromatin organization and transcriptional programs that support pluripotency transitions, germline specification, and genome stability in early developmental contexts. Its activity has been associated with modulation of DNA methylation-dependent pathways and coordination of reprogramming-related gene networks. Dysregulated DPPA3 expression patterns have been reported in studies of infertility, developmental abnormalities, and cancer-associated epigenetic remodeling, making it relevant for mechanistic investigations of cell state control.

STELLA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DPPA3 expression without altering the underlying DNA sequence.

STELLA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DPPA3 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 DPPA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous STELLA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DPPA3 locus and enabling the study of STELLA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of STELLA pathway restoration in tumor cells with silenced or reduced DPPA3 expression.

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