SHOX2 CRISPR Activation Plasmid (h): sc-402950-ACT

SHOX2 (short stature homeobox 2) encodes a homeobox transcription factor that regulates developmental gene programs controlling cellular identity, differentiation, and tissue patterning. In human cells, SHOX2 contributes to transcriptional networks that shape cardiogenesis and pacemaker lineage specification and can influence epithelial–mesenchymal features through downstream target modulation. Altered SHOX2 expression and promoter methylation have been linked to cancer-associated transcriptional reprogramming and to developmental phenotypes involving cardiac conduction and growth-related traits. As a DNA-binding regulator, SHOX2 provides a tractable node for interrogating gene regulatory circuitry, chromatin state effects, and lineage-restricted transcriptional control.

SHOX2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SHOX2 expression without altering the underlying DNA sequence.

SHOX2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SHOX2 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 SHOX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SHOX2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SHOX2 locus and enabling the study of SHOX2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SHOX2 pathway restoration in tumor cells with silenced or reduced SHOX2 expression.

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