GKLF/KLF4 CRISPR Activation Plasmid (m): sc-421298-ACT

Mouse Klf4 (GKLF/KLF4) encodes a zinc-finger transcription factor that regulates epithelial differentiation, cell-cycle control, and maintenance of cell identity by coordinating lineage-specific transcriptional programs. KLF4 modulates pathways linked to stemness and reprogramming, including integration with WNT/β-catenin, TGF-β/SMAD, and p53-dependent stress responses, and it influences barrier function and inflammatory signaling in epithelial tissues. Depending on cellular context, altered Klf4 expression perturbs proliferation–differentiation balance and can contribute to dysregulated tissue homeostasis relevant to oncogenic transformation, fibrosis, and wound-repair phenotypes. In mouse models, Klf4 is widely used to study somatic cell reprogramming, epithelial biology, and transcriptional networks governing pluripotency and differentiation.

GKLF/KLF4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Klf4 expression without altering the underlying DNA sequence.

GKLF/KLF4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Klf4 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 Klf4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GKLF/KLF4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Klf4 locus and enabling the study of GKLF/KLF4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GKLF/KLF4 pathway restoration in tumor cells with silenced or reduced Klf4 expression.

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