HN1 CRISPR Activation Plasmid (h): sc-412133-ACT

HN1 (hematological and neurological expressed 1) encodes a small, conserved protein implicated in regulating cell proliferation, differentiation, and cell-cycle progression across developmental and stress contexts. Although its molecular partners remain incompletely defined, HN1 expression has been linked to transcriptional programs associated with mitogenic signaling, epithelial–mesenchymal plasticity, and metabolic adaptation, suggesting roles in coordinating growth-state transitions. Dysregulated HN1 levels have been reported in multiple tumor types and are frequently evaluated as a biomarker-like correlate of aggressive cellular phenotypes, including altered invasion and therapy response signatures. These properties make HN1 a useful target for mechanistic studies of oncogenic signaling networks and lineage-state regulation in human cell models.

HN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HN1 expression without altering the underlying DNA sequence.

HN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HN1 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 HN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HN1 locus and enabling the study of HN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HN1 pathway restoration in tumor cells with silenced or reduced HN1 expression.

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