HoxB2 CRISPR Activation Plasmid (h): sc-404533-ACT

HOXB2 encodes the homeobox transcription factor HoxB2, a DNA-binding regulator that helps establish anterior–posterior patterning and segmental identity during embryonic development, particularly within hindbrain and neural crest-derived lineages. HoxB2 modulates gene expression programs controlling cell fate specification, differentiation, and morphogen responsiveness through HOX transcriptional networks and interactions with cofactors such as PBX and MEIS. Dysregulated HOXB2 expression has been associated with altered developmental signaling and aberrant transcriptional states in human disease contexts, including cancers where HOX gene programs can influence proliferation, migration, and lineage plasticity. As a transcriptional regulator, HoxB2 provides a tractable node for interrogating gene regulatory circuitry and developmental pathway reactivation in model systems.

HoxB2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXB2 expression without altering the underlying DNA sequence.

HoxB2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXB2 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 HOXB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxB2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXB2 locus and enabling the study of HoxB2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxB2 pathway restoration in tumor cells with silenced or reduced HOXB2 expression.

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