BTEB1 CRISPR Activation Plasmid (h): sc-402170-ACT

KLF9 (BTEB1) encodes a Krüppel-like transcription factor that binds GC-rich promoter elements to regulate context-dependent programs controlling cell-cycle progression, differentiation, and stress-responsive gene expression. In human cells, BTEB1 integrates signals from nuclear receptor and MAPK-associated pathways to shape transcriptional outputs linked to metabolic homeostasis and endocrine responsiveness. Altered KLF9 expression has been associated with dysregulated proliferation, inflammatory signaling, and lineage-specific remodeling, making it a useful node for studying transcriptional network perturbations. These properties support mechanistic research in chromatin regulation, stimulus-dependent transcription, and disease-relevant gene expression signatures.

BTEB1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KLF9 expression without altering the underlying DNA sequence.

BTEB1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KLF9 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 KLF9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTEB1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KLF9 locus and enabling the study of BTEB1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTEB1 pathway restoration in tumor cells with silenced or reduced KLF9 expression.

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