Chk2 CRISPR Activation Plasmid (h): sc-400438-ACT

CHEK2 encodes the human checkpoint kinase Chk2, a serine/threonine kinase activated downstream of ATM in response to DNA double-strand breaks. Chk2 propagates DNA damage signaling by phosphorylating effectors that regulate cell-cycle checkpoints, DNA repair, apoptosis, and replication stress responses, integrating with p53 and other genome surveillance networks. Through these pathways, CHEK2 helps preserve genomic stability and influences cellular outcomes following genotoxic stress. Altered CHEK2/Chk2 signaling is associated with defective checkpoint control and increased susceptibility to genome instability observed across multiple disease contexts, supporting its broad relevance in mechanistic studies.

Chk2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHEK2 expression without altering the underlying DNA sequence.

Chk2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHEK2 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 CHEK2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Chk2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHEK2 locus and enabling the study of Chk2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Chk2 pathway restoration in tumor cells with silenced or reduced CHEK2 expression.

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