KLF3 CRISPR Activation Plasmid (h): sc-402926-ACT

KLF3 (Krüppel-like factor 3) is a zinc-finger transcription factor that binds GC-rich regulatory elements to coordinate gene expression programs controlling erythroid maturation, adipogenesis, and broader hematopoietic lineage decisions. By functioning largely as a context-dependent transcriptional repressor through interactions with coregulator complexes, KLF3 modulates chromatin state and influences RNA polymerase II–dependent transcriptional output. KLF3-linked networks intersect with metabolic and immune regulatory pathways, including erythrocyte membrane and globin-related gene modules as well as inflammatory transcriptional circuits. Dysregulated KLF3 activity has been associated with altered differentiation states and transcriptional rewiring observed in hematologic and metabolic disease research, making it a useful target for mechanistic studies of cell identity and gene regulatory circuitry.

KLF3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLF3 expression without altering the underlying DNA sequence.

KLF3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLF3 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 KLF3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KLF3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLF3 locus and enabling the study of KLF3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KLF3 pathway restoration in tumor cells with silenced or reduced KLF3 expression.

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