HoxB5 CRISPR Activation Plasmid (m): sc-420914-ACT

Hoxb5 encodes the homeobox transcription factor HoxB5, a sequence-specific DNA-binding regulator that patterns the anterior–posterior axis and orchestrates regional identity during embryogenesis. In mouse, HoxB5 helps coordinate developmental gene regulatory networks that control progenitor cell specification, differentiation timing, and tissue morphogenesis, integrating with broader HOX cluster programs and chromatin-mediated transcriptional control. Its activity influences pathways governing organogenesis and hematopoietic and neural crest-associated processes, where altered HOX expression is frequently studied in the context of dysregulated differentiation and proliferative states. Because HOX factors sit high in regulatory hierarchies, Hoxb5 is widely used to probe developmental transcriptional circuitry and lineage programming mechanisms relevant to disease models.

HoxB5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hoxb5 expression without altering the underlying DNA sequence.

HoxB5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Hoxb5 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 Hoxb5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxB5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Hoxb5 locus and enabling the study of HoxB5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxB5 pathway restoration in tumor cells with silenced or reduced Hoxb5 expression.

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