Histone Deacetylase 3 (HDAC3) CRISPR Activation Plasmid (h): sc-400446-ACT

Histone deacetylase 3 (HDAC3) is a class I HDAC that forms core catalytic activity within the NCoR/SMRT co-repressor complexes to remove acetyl groups from histone tails and selected non-histone substrates, thereby shaping chromatin accessibility and transcriptional programs. Through epigenetic control of enhancer and promoter states, HDAC3 contributes to cell-cycle progression, DNA damage responses, and lineage-specific differentiation, and integrates signaling inputs that influence metabolic and inflammatory gene expression. Dysregulated HDAC3 activity or recruitment has been implicated in aberrant transcriptional repression across multiple disease contexts, including cancers and neurodevelopmental and immune-related disorders, where altered chromatin acetylation is a recurring molecular feature. As a nodal epigenetic regulator, HDAC3 is frequently studied for its impact on transcriptional networks, chromatin remodeling, and context-dependent control of cellular identity.

Histone Deacetylase 3 (HDAC3) CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HDAC3 expression without altering the underlying DNA sequence.

Histone Deacetylase 3 (HDAC3) CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HDAC3 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 HDAC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Histone Deacetylase 3 (HDAC3) expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HDAC3 locus and enabling the study of Histone Deacetylase 3 (HDAC3)-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Histone Deacetylase 3 (HDAC3) pathway restoration in tumor cells with silenced or reduced HDAC3 expression.

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