BTEB2 CRISPR Activation Plasmid (h): sc-401680-ACT

KLF5 (BTEB2) encodes a Kruppel-like zinc finger transcription factor that binds GC-rich promoter elements to regulate programs controlling cell-cycle progression, epithelial differentiation, and tissue remodeling. It integrates signals from MAPK/ERK and TGF-β/SMAD pathways and coordinates transcriptional responses that influence proliferation, migration, and extracellular matrix dynamics. Dysregulated KLF5 activity has been linked to oncogenic and inflammatory phenotypes in multiple tissues, making it a widely used node for studying transcriptional control of growth and stress responses. In human cell models, perturbation of BTEB2 helps dissect context-dependent gene regulatory networks and pathway cross-talk underlying disease-associated cellular states.

BTEB2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLF5 expression without altering the underlying DNA sequence.

BTEB2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLF5 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 KLF5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTEB2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLF5 locus and enabling the study of BTEB2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTEB2 pathway restoration in tumor cells with silenced or reduced KLF5 expression.

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