CASZ1 CRISPR Activation Plasmid (h): sc-404756-ACT

CASZ1 (castor zinc finger 1) encodes a human transcription factor that binds DNA through multiple C2H2 zinc-finger domains to regulate gene expression programs controlling cell fate specification, differentiation, and tissue morphogenesis. CASZ1 activity has been linked to developmental signaling networks that shape neuronal and cardiovascular lineages, including pathways influencing cell-cycle progression and cytoskeletal organization. Dysregulated CASZ1 expression or altered regulatory circuitry is associated with disease-relevant phenotypes such as impaired differentiation states and aberrant proliferation, with reported relevance in neuroblastoma biology and vascular development. As an endogenous transcriptional regulator, CASZ1 is widely studied for how lineage-restricted transcriptional programs interface with chromatin regulation and developmental gene networks.

CASZ1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CASZ1 expression without altering the underlying DNA sequence.

CASZ1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CASZ1 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 CASZ1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CASZ1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CASZ1 locus and enabling the study of CASZ1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CASZ1 pathway restoration in tumor cells with silenced or reduced CASZ1 expression.

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