HoxD10 CRISPR Activation Plasmid (h): sc-403328-ACT

HOXD10 encodes the homeobox transcription factor HoxD10, a key regulator of anterior–posterior patterning and limb morphogenesis that helps establish cell identity through sequence-specific DNA binding and transcriptional control. In differentiated tissues, HOXD10 contributes to programs governing cell migration, adhesion, and extracellular matrix remodeling, interfacing with developmental transcriptional networks and signaling pathways that shape lineage commitment. Altered HOXD10 expression or regulatory disruption has been associated with oncogenic phenotypes, including changes in invasiveness and metastatic potential, reflecting its role in controlling gene sets linked to epithelial–mesenchymal dynamics. As a developmental regulator with context-dependent activity, HOXD10 is frequently used to study transcriptional circuitry, chromatin regulation, and disease-relevant gene expression states in human cell models.

HoxD10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXD10 expression without altering the underlying DNA sequence.

HoxD10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXD10 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 HOXD10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxD10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXD10 locus and enabling the study of HoxD10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxD10 pathway restoration in tumor cells with silenced or reduced HOXD10 expression.

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