HMGI-C CRISPR Activation Plasmid (m): sc-420880-ACT

Hmga2 encodes the architectural chromatin protein HMGI-C, a non-histone AT-hook factor that binds AT-rich DNA and reshapes local chromatin to modulate transcriptional programs. In mouse cells, HMGI-C contributes to regulation of proliferation, differentiation, and developmental timing by influencing enhancer–promoter communication and transcription factor occupancy across the genome. It is frequently linked to stem/progenitor-like states and epithelial–mesenchymal transition-associated gene expression, intersecting with growth factor–responsive pathways that coordinate cell cycle progression and lineage commitment. Dysregulated HMGA2/HMGI-C expression or rearrangement is associated with aberrant growth and oncogenic transcriptional signatures, making it a useful node for mechanistic studies of chromatin-driven gene regulation.

HMGI-C CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hmga2 expression without altering the underlying DNA sequence.

HMGI-C CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Hmga2 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 Hmga2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HMGI-C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Hmga2 locus and enabling the study of HMGI-C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HMGI-C pathway restoration in tumor cells with silenced or reduced Hmga2 expression.

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