ULK2 CRISPR Activation Plasmid (h): sc-402558-ACT

ULK2 (UNC-51 like autophagy activating kinase 2) encodes a serine/threonine kinase that functions as a core initiator of macroautophagy, coordinating early autophagosome formation in response to nutrient and energy cues. ULK2 operates within the ULK complex and integrates upstream inputs from mTORC1 and AMPK signaling to modulate phosphorylation events that link metabolic stress to autophagic flux. Through its role in proteostasis, organelle quality control, and cellular adaptation to starvation, ULK2 influences processes such as mitochondrial homeostasis and stress resilience. Dysregulation of ULK2-associated autophagy programs has been investigated in contexts relevant to neurodegeneration, cancer cell metabolism, and inflammatory signaling.

ULK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ULK2 expression without altering the underlying DNA sequence.

ULK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ULK2 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 ULK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ULK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ULK2 locus and enabling the study of ULK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ULK2 pathway restoration in tumor cells with silenced or reduced ULK2 expression.

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