HoxA10 CRISPR Activation Plasmid (h): sc-401463-ACT

HOXA10 encodes the homeobox transcription factor HoxA10, a sequence-specific DNA-binding regulator that patterns embryonic development and maintains adult tissue identity. In human cells, HoxA10 contributes to anterior–posterior regionalization, organogenesis, and lineage specification by coordinating transcriptional programs that intersect with developmental signaling networks such as retinoic acid-responsive pathways and hormone-regulated gene expression. Its activity is particularly relevant to uterine/endometrial biology and hematopoietic differentiation, where altered expression can shift proliferation and differentiation states. Dysregulated HOXA10 expression has been associated with developmental abnormalities and has been studied in the context of reproductive disorders and hematologic malignancy-related transcriptional rewiring.

HoxA10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HOXA10 expression without altering the underlying DNA sequence.

HoxA10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXA10 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 HOXA10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxA10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXA10 locus and enabling the study of HoxA10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxA10 pathway restoration in tumor cells with silenced or reduced HOXA10 expression.

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