HoxA1 CRISPR Activation Plasmid (h): sc-403158-ACT

HOXA1 encodes the human homeobox transcription factor HoxA1, a sequence-specific DNA-binding regulator that patterns the early anterior–posterior axis and guides hindbrain and craniofacial development. Through control of lineage specification programs, HOXA1 influences transcriptional networks that intersect with retinoic acid–responsive signaling and broader chromatin-regulated differentiation processes. Dysregulated HOXA1 expression has been associated with altered developmental gene expression states and has been investigated in contexts of congenital malformations and oncogenic transcriptional reprogramming. As a developmental regulator, HOXA1 is widely used as a marker and mechanistic node for studying cell fate transitions and morphogen-driven patterning.

HoxA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXA1 expression without altering the underlying DNA sequence.

HoxA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXA1 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 HOXA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXA1 locus and enabling the study of HoxA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxA1 pathway restoration in tumor cells with silenced or reduced HOXA1 expression.

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