PASK CRISPR Activation Plasmid (h): sc-416953-ACT

Per-Arnt-Sim (PAS) domain-containing serine/threonine-protein kinase (PASK) functions as a nutrient- and energy-sensing kinase that links metabolic cues to gene expression and cell fate decisions. In human cells, PASK integrates signals related to glucose availability and mitochondrial function and modulates downstream transcriptional programs involved in carbohydrate and lipid metabolism, including pathways connected to pancreatic endocrine differentiation and insulin-responsive processes. PASK activity has been studied in the context of AMPK- and mTOR-adjacent signaling networks, where it can influence anabolic–catabolic balance and stress-adaptive responses. Dysregulated PASK expression or signaling has been associated with metabolic phenotypes relevant to diabetes and obesity research, and has also been explored for its broader roles in proliferation and cellular adaptation.

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

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

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