FOG CRISPR Activation Plasmid (h): sc-402620-ACT

Human ZFPM1 encodes Friend of GATA-1 (FOG), a multi–zinc finger transcriptional cofactor that binds GATA family proteins to modulate lineage-specific gene programs. FOG coordinates activating and repressive chromatin complexes, including NuRD/HDAC-associated machinery, to regulate erythroid and megakaryocytic differentiation as well as broader hematopoietic development. Through its control of transcriptional networks governing cell fate commitment, ZFPM1 is frequently studied in pathways that shape hematopoiesis, platelet biology, and developmental gene regulation. Dysregulated ZFPM1/FOG activity and altered GATA cofactor balance are relevant to aberrant hematopoietic differentiation and related research models of blood and developmental disorders.

FOG CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ZFPM1 expression without altering the underlying DNA sequence.

FOG CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ZFPM1 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 ZFPM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOG expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ZFPM1 locus and enabling the study of FOG-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOG pathway restoration in tumor cells with silenced or reduced ZFPM1 expression.

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