Nanog CRISPR Activation Plasmid (m): sc-428155-ACT

Mouse Nanog encodes a homeobox transcription factor that serves as a central regulator of pluripotency and self-renewal in the preimplantation embryo and embryonic stem cells. NANOG cooperates with OCT4 and SOX2 to establish core transcriptional circuitry, while interacting with chromatin remodeling and epigenetic control pathways that govern lineage commitment and repression of differentiation programs. Altered NANOG expression is widely used as a marker of stem-like cellular states and is implicated in mechanisms that support aberrant self-renewal, epithelial–mesenchymal transition, and resistance to differentiation cues in disease-relevant models. These properties make Nanog a key target for investigating developmental gene regulatory networks, cell fate transitions, and transcription factor–driven reprogramming.

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

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

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