SIK2 CRISPR Activation Plasmid (h): sc-406982-ACT

Salt-inducible kinase 2 (SIK2) is a serine/threonine kinase in the AMPK-related kinase family that coordinates cellular energy sensing with transcriptional programs. SIK2 regulates phosphorylation-dependent control of CREB coactivators (CRTCs) and class IIa HDACs, thereby influencing cAMP/CREB signaling, metabolic gene expression, and stress-responsive pathways. In human cells, SIK2 activity has been linked to adipocyte differentiation and lipid/glucose homeostasis, and it also contributes to cell-cycle progression and cytoskeletal dynamics through modulation of kinase signaling networks. Dysregulated SIK2 expression or signaling is investigated in cancer biology and metabolic disease models, where altered transcriptional control and proliferation-associated pathways are key mechanistic readouts.

SIK2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SIK2 expression without altering the underlying DNA sequence.

SIK2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SIK2 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 SIK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SIK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SIK2 locus and enabling the study of SIK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SIK2 pathway restoration in tumor cells with silenced or reduced SIK2 expression.

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