HoxB13 CRISPR Activation Plasmid (h): sc-401878-ACT

HOXB13 encodes the homeobox transcription factor HoxB13, a sequence-specific DNA-binding regulator that helps establish positional identity and lineage programs during embryonic development and tissue differentiation. By controlling transcriptional networks that influence cell fate decisions, epithelial maturation, and organ patterning, HoxB13 contributes to context-dependent regulation of proliferation and differentiation. Dysregulated HOXB13 expression or function has been linked to altered gene expression programs in hormone-responsive tissues, with particular relevance to prostate biology and cancer-associated transcriptional states. As a nuclear transcription factor, HoxB13 is widely studied for its roles in developmental pathways and disease-relevant transcriptional circuitry.

HoxB13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXB13 expression without altering the underlying DNA sequence.

HoxB13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXB13 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 HOXB13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxB13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXB13 locus and enabling the study of HoxB13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxB13 pathway restoration in tumor cells with silenced or reduced HOXB13 expression.

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