HoxB9 CRISPR Activation Plasmid (h): sc-401770-ACT

HOXB9 encodes the homeobox transcription factor HoxB9, a sequence-specific DNA-binding protein that regulates anterior–posterior patterning, tissue morphogenesis, and cell fate decisions during development. In human cells, HOXB9 contributes to transcriptional programs that influence differentiation and proliferation, integrating with broader developmental gene regulatory networks such as HOX-cofactor interactions and chromatin-mediated control of lineage-specific enhancers. Dysregulated HOXB9 expression has been associated with altered epithelial–mesenchymal dynamics, angiogenic signaling, and invasive phenotypes in multiple cancer contexts, making it a useful node for studying oncogenic transcriptional rewiring. As a nuclear regulator, HoxB9 is frequently examined for its impact on gene expression circuitry governing migration, extracellular matrix remodeling, and stem-like states.

HoxB9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXB9 expression without altering the underlying DNA sequence.

HoxB9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXB9 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 HOXB9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxB9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXB9 locus and enabling the study of HoxB9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxB9 pathway restoration in tumor cells with silenced or reduced HOXB9 expression.

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