MOF CRISPR Activation Plasmid (h): sc-401891-ACT

KAT8 encodes MOF (MYST1), a MYST-family histone acetyltransferase that catalyzes acetylation of histone H4 lysine 16 (H4K16ac), a key chromatin mark linked to open chromatin and transcriptional competence. MOF functions within multiprotein complexes to regulate promoter and enhancer activity, coordinate transcriptional elongation, and modulate chromatin architecture during DNA replication and repair. Through these epigenetic roles, MOF influences genome stability, cell-cycle progression, and stress responses, with altered KAT8/MOF activity and H4K16ac patterns frequently studied in contexts of oncogenic transcription programs and dysregulated DNA damage signaling. Its central position in chromatin regulation also makes KAT8 a useful node for interrogating crosstalk between acetylation-dependent chromatin remodeling and signaling pathways controlling proliferation and differentiation.

MOF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KAT8 expression without altering the underlying DNA sequence.

MOF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KAT8 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 KAT8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MOF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KAT8 locus and enabling the study of MOF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MOF pathway restoration in tumor cells with silenced or reduced KAT8 expression.

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