HIG2 CRISPR Activation Plasmid (h): sc-403510-ACT

Human HILPDA encodes hypoxia-inducible lipid droplet–associated protein (HIG2), a small stress-response factor that localizes to lipid droplets and interfaces with metabolic remodeling under low oxygen and nutrient limitation. HIG2 is induced downstream of HIF signaling and contributes to regulation of intracellular lipid storage and utilization, influencing lipolysis control and fatty acid handling during hypoxic adaptation. Through these processes, HILPDA/HIG2 is relevant to pathways linking hypoxia, lipid metabolism, and inflammatory or metabolic stress responses. Dysregulated expression has been associated with conditions characterized by altered oxygen tension and lipid reprogramming, supporting its use as a mechanistic node for studying disease-relevant metabolic phenotypes.

HIG2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HILPDA expression without altering the underlying DNA sequence.

HIG2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HILPDA 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 HILPDA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HIG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HILPDA locus and enabling the study of HIG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HIG2 pathway restoration in tumor cells with silenced or reduced HILPDA expression.

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