PLZF CRISPR Activation Plasmid (h): sc-400354-ACT

ZBTB16 encodes PLZF, a BTB/POZ-domain zinc finger transcription factor that governs cell fate decisions by coordinating chromatin-associated transcriptional repression and activation programs. In human hematopoiesis and immune development, PLZF helps regulate progenitor maintenance and lineage specification, including effects on innate-like lymphocyte differentiation and cytokine-responsive gene expression. PLZF-dependent networks intersect with epigenetic control of proliferation and differentiation, linking this factor to developmental regulation and cellular maturation pathways. Dysregulated ZBTB16/PLZF expression or rearrangements have been associated with altered transcriptional programs in hematologic disease contexts, making it a useful node for studying transcription factor–driven phenotypes.

PLZF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZBTB16 expression without altering the underlying DNA sequence.

PLZF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZBTB16 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 ZBTB16 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLZF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZBTB16 locus and enabling the study of PLZF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLZF pathway restoration in tumor cells with silenced or reduced ZBTB16 expression.

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