GCN5 CRISPR Activation Plasmid (m): sc-420512-ACT

Mouse Kat2a encodes GCN5, a lysine acetyltransferase that acetylates histone H3 (including H3K9 and H3K14) to promote chromatin accessibility and transcriptional activation. As a core component of multi-subunit coactivator complexes such as SAGA and ATAC, GCN5 integrates signaling inputs with RNA polymerase II–dependent gene expression programs that control cell cycle progression, differentiation, and stress responses. Kat2a activity influences enhancer and promoter dynamics, DNA damage response, and metabolic gene regulation through coordinated histone acetylation and transcription factor coactivation. Dysregulated GCN5-dependent epigenetic control has been associated with aberrant proliferation and lineage specification, making Kat2a a useful node for modeling chromatin-driven mechanisms in disease-relevant cellular states.

GCN5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kat2a expression without altering the underlying DNA sequence.

GCN5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kat2a 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 Kat2a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GCN5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kat2a locus and enabling the study of GCN5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GCN5 pathway restoration in tumor cells with silenced or reduced Kat2a expression.

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