PASK CRISPR Activation Plasmid (m): sc-435323-ACT

Mouse Pask encodes PAS domain–containing serine/threonine-protein kinase (PASK), a metabolic sensor that integrates nutrient and energy cues with transcriptional and post-translational control. PASK participates in signaling networks linked to glucose homeostasis, mitochondrial function, and regulation of protein synthesis, with reported connections to mTOR-related processes and stress-responsive adaptation. In diverse cell types, PASK activity can influence proliferation and differentiation programs by modulating downstream effector phosphorylation and transcriptional outputs. Dysregulated PASK signaling has been associated with metabolic and endocrine phenotypes, making it relevant for mechanistic studies of insulin signaling, lipid handling, and cellular energy balance.

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

PASK CRISPR Activation Plasmid (m) 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.