Krs-1 CRISPR Activation Plasmid (h): sc-416738-ACT

Human STK3 encodes the serine/threonine kinase Krs-1 (also known as MST2), a core component of the Hippo signaling cascade that constrains cell proliferation and supports contact inhibition by phosphorylating downstream kinases that regulate YAP/TAZ-dependent transcription. Krs-1 also interfaces with stress-responsive and apoptotic processes through kinase-driven signaling outputs that influence cytoskeletal dynamics, cell polarity, and survival decisions. Dysregulation of STK3/Hippo pathway activity is frequently studied in the context of altered growth control, epithelial–mesenchymal transitions, and tumor-associated signaling rewiring. These features make STK3 a useful node for dissecting pathway crosstalk and transcriptional programs governing tissue homeostasis in human cell models.

Krs-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STK3 expression without altering the underlying DNA sequence.

Krs-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the STK3 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 STK3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Krs-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native STK3 locus and enabling the study of Krs-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Krs-1 pathway restoration in tumor cells with silenced or reduced STK3 expression.

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