Histone Deacetylase 4 (HDAC4) CRISPR Activation Plasmid (h): sc-400388-ACT

HDAC4 encodes Histone Deacetylase 4, a class IIa HDAC that functions as a signal-responsive chromatin regulator linking extracellular cues to transcriptional programs. HDAC4 shuttles between cytoplasm and nucleus in response to calcium/calmodulin-dependent kinases and other signaling inputs, where it modulates histone acetylation states and represses transcription in part through interactions with transcription factors such as MEF2. Through these mechanisms, HDAC4 participates in control of cell differentiation, neuronal and muscle gene expression, stress responses, and epigenetic remodeling. Dysregulated HDAC4 activity or localization has been implicated in pathological transcriptional states relevant to neurodevelopmental and neurodegenerative processes, cardiac remodeling, and cancer-associated gene expression changes.

Histone Deacetylase 4 (HDAC4) CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HDAC4 expression without altering the underlying DNA sequence.

Histone Deacetylase 4 (HDAC4) CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HDAC4 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 HDAC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Histone Deacetylase 4 (HDAC4) expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HDAC4 locus and enabling the study of Histone Deacetylase 4 (HDAC4)-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Histone Deacetylase 4 (HDAC4) pathway restoration in tumor cells with silenced or reduced HDAC4 expression.

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