HNF-1β CRISPR Activation Plasmid (h): sc-400537-ACT

HNF1B encodes the transcription factor HNF‑1β, a homeodomain-containing regulator essential for epithelial differentiation and organ development, particularly in kidney, pancreas, and liver lineages. It controls gene networks involved in tubular transport, cell polarity, and metabolic homeostasis, interfacing with pathways that shape nephron segmentation and ductal epithelial identity. Altered HNF1B expression or function is linked to renal developmental abnormalities, cystic and tubulointerstitial phenotypes, and dysregulated glucose metabolism consistent with monogenic diabetes syndromes. In cancer biology, HNF‑1β is frequently studied as a lineage and differentiation factor in epithelial tumors, where it can influence transcriptional programs connected to proliferation and stress responses.

HNF-1β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HNF1B expression without altering the underlying DNA sequence.

HNF-1β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HNF1B 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 HNF1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HNF-1β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HNF1B locus and enabling the study of HNF-1β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HNF-1β pathway restoration in tumor cells with silenced or reduced HNF1B expression.

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