HoxA5 CRISPR Activation Plasmid (h): sc-402499-ACT

HOXA5 encodes the homeobox transcription factor HoxA5, a sequence-specific DNA-binding regulator that helps establish anterior–posterior patterning and segment identity during human development. In differentiated tissues, HoxA5 contributes to programs controlling cell fate commitment, epithelial differentiation, and tissue morphogenesis by coordinating transcriptional networks with other HOX cofactors. Dysregulated HOXA5 expression has been associated with altered proliferation, invasion, and lineage state changes in multiple disease contexts, making it a useful node for studying developmental reprogramming and transcriptional control. As a regulator of gene expression, HoxA5 is often examined in pathways linked to differentiation, extracellular matrix remodeling, and context-dependent growth control.

HoxA5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXA5 expression without altering the underlying DNA sequence.

HoxA5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXA5 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 HOXA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxA5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXA5 locus and enabling the study of HoxA5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxA5 pathway restoration in tumor cells with silenced or reduced HOXA5 expression.

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