CIDE-A CRISPR Activation Plasmid (h): sc-405086-ACT

CIDEA (cell death–inducing DFFA-like effector A) encodes the lipid droplet–associated protein CIDE-A, a key regulator of neutral lipid storage and adipocyte metabolism. In human adipose and brown/beige fat–related programs, CIDE-A supports lipid droplet growth and remodeling and influences triglyceride turnover, coupling lipid storage dynamics to mitochondrial activity and thermogenic gene networks. CIDEA expression and function are tightly linked to energy homeostasis pathways, including adipogenesis and lipid handling processes, and have been associated with metabolic phenotypes such as obesity, insulin resistance, and fatty liver–related lipid dysregulation. As a tissue- and state-dependent metabolic effector, CIDE-A is commonly studied for its roles in adipocyte differentiation, lipid droplet biology, and stress responses in metabolically active cells.

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

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

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