NPRL2 CRISPR Activation Plasmid (h): sc-402875-ACT

Human NPRL2 encodes a core component of the GATOR1 complex, a key negative regulator of mTORC1 signaling that links amino acid availability to cellular growth control. By promoting restraint of mTORC1 activity, NPRL2 supports nutrient-stress responses, autophagy regulation, and metabolic homeostasis, influencing protein synthesis and cell cycle progression. Altered NPRL2 expression or function has been associated with dysregulated mTOR pathway activity observed in diverse contexts of aberrant proliferation and stress adaptation, making it relevant for mechanistic studies of growth control. NPRL2 is also used as a node to interrogate nutrient-sensing circuitry and downstream transcriptional programs that shape cellular survival and biosynthetic capacity.

NPRL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPRL2 expression without altering the underlying DNA sequence.

NPRL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPRL2 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 NPRL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPRL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPRL2 locus and enabling the study of NPRL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPRL2 pathway restoration in tumor cells with silenced or reduced NPRL2 expression.

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