Pax CRISPR Activation Plasmid (h): sc-402171-ACT

PAX2 encodes the paired box transcription factor Pax2, a sequence-specific DNA-binding regulator essential for embryonic patterning and lineage specification, particularly in the developing kidney, urogenital tract, eye, and central nervous system. Pax2 coordinates gene programs controlling epithelial differentiation, cell survival, and morphogenesis through transcriptional networks that intersect with developmental signaling pathways such as WNT, FGF, and NOTCH. Dysregulated PAX2 expression or mutation has been associated with congenital anomalies of the kidney and urinary tract and ocular developmental disorders, and altered Pax2 activity is also studied in contexts of aberrant proliferation and lineage plasticity. As a developmental regulator, PAX2 is widely used as a marker and functional node in nephrogenesis models, organoid systems, and studies of epithelial identity.

Pax-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAX2 expression without altering the underlying DNA sequence.

Pax-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAX2 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 PAX2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pax-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAX2 locus and enabling the study of Pax-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pax-2 pathway restoration in tumor cells with silenced or reduced PAX2 expression.

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