Six1 CRISPR Activation Plasmid (h): sc-402063-ACT

SIX1 encodes Six1, a homeobox transcription factor that regulates embryonic development programs including organogenesis, myogenesis, and sensory placode formation by coordinating lineage-specific gene expression networks. Six1 functions within transcriptional regulatory circuits that interface with developmental signaling pathways such as WNT, FGF, and TGF-β, shaping cell fate decisions, proliferation, and differentiation. In adult tissues, dysregulated SIX1 expression can promote aberrant transcriptional states linked to epithelial–mesenchymal transition, invasion-associated programs, and altered cell cycle control. As a result, SIX1 is widely studied as a driver of developmental gene reactivation in cancer biology and as a determinant of tissue-specific differentiation phenotypes.

Six1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SIX1 expression without altering the underlying DNA sequence.

Six1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SIX1 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 SIX1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Six1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SIX1 locus and enabling the study of Six1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Six1 pathway restoration in tumor cells with silenced or reduced SIX1 expression.

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