HoxA11 CRISPR Activation Plasmid (h2): sc-405264-ACT-2

Human HOXA11 encodes the homeobox transcription factor HoxA11, a DNA-binding regulator that helps establish anterior–posterior patterning and regional identity during embryogenesis and contributes to differentiation programs in mesenchymal and reproductive tract lineages. HoxA11 modulates gene expression through homeobox motif recognition and cofactor-dependent transcriptional control, interfacing with developmental signaling networks that govern morphogenesis, tissue remodeling, and cell fate decisions. Dysregulated HOXA11 expression or epigenetic alteration has been associated with congenital malformations and cancers in which aberrant HOX gene programs support altered differentiation and proliferation states. HOXA11-targeted gene editing and functional genomics assays are used to map cis-regulatory elements, define downstream transcriptional circuits, and interrogate developmental and disease-relevant phenotypes in human cell models.

HoxA11 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous HOXA11 expression without altering the underlying DNA sequence.

HoxA11 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HOXA11 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 HOXA11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HoxA11 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HOXA11 locus and enabling the study of HoxA11-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HoxA11 pathway restoration in tumor cells with silenced or reduced HOXA11 expression.

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