SIK1 CRISPR Activation Plasmid (h): sc-401769-ACT

Salt-inducible kinase 1 (SIK1) is a serine/threonine kinase in the AMPK-related kinase family that transduces cAMP/PKA-dependent signals to regulate transcriptional programs controlling metabolism, circadian signaling, and stress responses. SIK1 modulates CREB/CRTC activity and contributes to phosphorylation-dependent control of transcriptional coactivators, linking extracellular cues to gene expression changes in multiple cell types. Through these pathways, SIK1 influences cellular energy homeostasis, differentiation, and adaptive responses to environmental stimuli. Dysregulated SIK1 signaling has been associated with altered metabolic regulation and aberrant transcriptional control in disease-relevant contexts, supporting its use in mechanistic studies of signaling-to-transcription coupling.

SIK1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SIK1 expression without altering the underlying DNA sequence.

SIK1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SIK1 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 SIK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SIK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SIK1 locus and enabling the study of SIK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SIK1 pathway restoration in tumor cells with silenced or reduced SIK1 expression.

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