KLF6 CRISPR Activation Plasmid (h): sc-400888-ACT

KLF6 (Krüppel-like factor 6) is a human zinc finger transcription factor that regulates gene programs controlling cell-cycle progression, differentiation, and cellular stress responses. It modulates transcriptional networks linked to TGF-β signaling and MAPK-dependent pathways, influencing expression of targets involved in growth restraint and extracellular matrix remodeling. KLF6 activity contributes to maintenance of tissue homeostasis and is frequently implicated in dysregulated transcriptional states observed in cancer biology and fibrotic processes. As a context-dependent regulator, KLF6 serves as a useful node for dissecting transcriptional circuitry and genotype-to-phenotype relationships across diverse cell models.

KLF6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLF6 expression without altering the underlying DNA sequence.

KLF6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLF6 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 KLF6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KLF6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLF6 locus and enabling the study of KLF6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KLF6 pathway restoration in tumor cells with silenced or reduced KLF6 expression.

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