ZBP-89 CRISPR Activation Plasmid (h): sc-404898-ACT

Human ZNF148 encodes the Krüppel-like zinc finger transcription factor ZBP-89, a GC-rich promoter-binding protein that modulates RNA polymerase II transcriptional programs controlling cell-cycle progression, differentiation, and cellular stress responses. ZBP-89 can act as an activator or repressor depending on promoter context and cofactor recruitment, influencing chromatin state and transcriptional initiation at genes involved in growth control and apoptosis. Through its roles in transcriptional regulation, ZBP-89 intersects with pathways governing epithelial homeostasis and lineage specification, making it relevant to studies of dysregulated proliferation and tumor-associated transcriptional networks. Altered ZNF148 activity has been examined in the context of cancer biology and inflammatory signaling, supporting its use as a mechanistic node for gene regulatory and promoter function research.

ZBP-89 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZNF148 expression without altering the underlying DNA sequence.

ZBP-89 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZNF148 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 ZNF148 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZBP-89 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZNF148 locus and enabling the study of ZBP-89-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZBP-89 pathway restoration in tumor cells with silenced or reduced ZNF148 expression.

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