HoxA1 CRISPR Activation Plasmid (m): sc-420899-ACT

Hoxa1 encodes the homeobox transcription factor HoxA1, an early developmental regulator that helps establish anterior–posterior patterning and positional identity during embryogenesis. HoxA1 functions in transcriptional programs that coordinate hindbrain segmentation, neural crest migration, and organogenesis, integrating with broader HOX network regulation and signaling inputs such as retinoic acid–dependent gene expression. In mouse models, altered Hoxa1 activity disrupts normal morphogenesis and cell fate decisions, making it relevant for studying developmental defects and gene regulatory circuitry. As a sequence-specific DNA-binding factor, HoxA1 provides a tractable node for interrogating how developmental transcription factors reshape chromatin and downstream pathway outputs.

HoxA1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hoxa1 expression without altering the underlying DNA sequence.

HoxA1 CRISPR Activation Plasmid (m) 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.