Miz-1 CRISPR Activation Plasmid (h): sc-402110-ACT

ZBTB17 encodes the transcription factor Miz-1, a BTB/POZ zinc-finger protein that binds initiator elements and coordinates context-dependent transcriptional programs controlling cell-cycle progression, differentiation, and stress responses. Miz-1 integrates signaling with transcriptional regulation through interactions with cofactors such as MYC and chromatin-modifying complexes, thereby modulating expression of genes including CDKN1A/p21 and other checkpoint-associated regulators. Through these networks, Miz-1 influences proliferation–apoptosis balance and lineage-specific gene expression, making ZBTB17 a useful node for studying transcriptional control in oncogenic and developmental contexts. Dysregulation of Miz-1–centered transcriptional circuits has been linked to altered cell-cycle checkpoints and aberrant growth phenotypes relevant to cancer biology and hematopoietic regulation.

Miz-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZBTB17 expression without altering the underlying DNA sequence.

Miz-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZBTB17 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 ZBTB17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Miz-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZBTB17 locus and enabling the study of Miz-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Miz-1 pathway restoration in tumor cells with silenced or reduced ZBTB17 expression.

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