ULK1 CRISPR Activation Plasmid (h): sc-400516-ACT

ULK1 (UNC-51 like autophagy activating kinase 1) is a serine/threonine kinase that initiates macroautophagy by integrating nutrient and energy signals. It functions at the core of the ULK1 complex to promote phagophore formation and coordinates downstream autophagy machinery in response to mTORC1 inhibition and AMPK activation. Through regulation of autophagic flux, ULK1 influences mitochondrial homeostasis, proteostasis, and cellular adaptation to metabolic stress. Dysregulated ULK1 signaling and autophagy control are implicated in contexts such as neurodegeneration, infection biology, and cancer cell stress responses, where altered catabolic capacity can reshape survival pathways.

ULK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ULK1 expression without altering the underlying DNA sequence.

ULK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ULK1 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 ULK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ULK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ULK1 locus and enabling the study of ULK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ULK1 pathway restoration in tumor cells with silenced or reduced ULK1 expression.

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