LRF/Pokemon CRISPR Activation Plasmid (h): sc-402016-ACT

ZBTB7A encodes the transcription factor LRF/Pokemon, a BTB/POZ zinc-finger protein that regulates gene expression programs controlling cell fate, proliferation, and differentiation. LRF/Pokemon participates in chromatin-associated transcriptional repression and activation, influencing lineage decisions in hematopoietic and other developmental contexts while intersecting with pathways governing cell-cycle control and stress responses. Dysregulated ZBTB7A activity has been linked to altered transcriptional networks in cancer biology and to perturbations in erythroid and immune cell development, making it a useful node for studying oncogenic transcription factor circuitry. Investigating ZBTB7A function supports mechanistic work on promoter/enhancer regulation, epigenetic state transitions, and context-dependent transcriptional rewiring.

LRF/Pokemon CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZBTB7A expression without altering the underlying DNA sequence.

LRF/Pokemon CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZBTB7A 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 ZBTB7A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRF/Pokemon expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZBTB7A locus and enabling the study of LRF/Pokemon-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRF/Pokemon pathway restoration in tumor cells with silenced or reduced ZBTB7A expression.

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