Histone Deacetylase 6 (HDAC6) CRISPR Activation Plasmid (h): sc-400314-ACT

HDAC6 encodes Histone Deacetylase 6, a predominantly cytoplasmic deacetylase that regulates acetylation of non-histone substrates including α-tubulin, cortactin, and HSP90, thereby influencing microtubule dynamics, cell motility, and chaperone-dependent proteostasis. Through control of ubiquitin-dependent aggresome formation and autophagic clearance, HDAC6 integrates cytoskeletal remodeling with stress responses to misfolded proteins. HDAC6 also modulates signaling in pathways linked to innate immunity and inflammatory transcriptional programs, including effects on NF-κB-regulated outputs. Dysregulated HDAC6 activity or expression has been associated with cancer cell phenotypes, neurodegeneration-related proteotoxic stress, and immune-mediated disease mechanisms, making it a useful node for mechanistic studies.

Histone Deacetylase 6 (HDAC6) CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HDAC6 expression without altering the underlying DNA sequence.

Histone Deacetylase 6 (HDAC6) CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HDAC6 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 HDAC6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Histone Deacetylase 6 (HDAC6) expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HDAC6 locus and enabling the study of Histone Deacetylase 6 (HDAC6)-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Histone Deacetylase 6 (HDAC6) pathway restoration in tumor cells with silenced or reduced HDAC6 expression.

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