LKB1 CRISPR Activation Plasmid (m): sc-423192-ACT

Mouse Stk11 encodes the serine/threonine kinase LKB1, a master regulator of cellular energy homeostasis that phosphorylates and activates AMPK family kinases. Through AMPK and related pathways, LKB1 coordinates metabolic stress responses, mitochondrial function, autophagy, cell polarity, and cell-cycle control, integrating nutrient sensing with growth regulation via mTOR signaling. Disruption or dysregulation of LKB1 signaling is broadly linked to altered metabolism and loss of polarity in disease-relevant contexts, making Stk11 a key node for studying stress adaptation and tissue homeostasis. In mouse models, Stk11/LKB1 modulation supports mechanistic interrogation of metabolic rewiring and signaling cross-talk in diverse cell types.

LKB1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Stk11 expression without altering the underlying DNA sequence.

LKB1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Stk11 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 Stk11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LKB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Stk11 locus and enabling the study of LKB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LKB1 pathway restoration in tumor cells with silenced or reduced Stk11 expression.

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