MafB CRISPR Activation Plasmid (h): sc-400554-ACT

MAFB encodes MafB, a large Maf bZIP transcription factor that binds Maf recognition elements to control lineage-specific transcription programs. In human tissues it is a key regulator of myeloid and macrophage differentiation, modulation of inflammatory gene expression, and maintenance of specialized cell states including renal podocyte identity and pancreatic islet function. MafB integrates signals from developmental and stress-responsive pathways to coordinate terminal differentiation, cellular maturation, and tissue homeostasis. Dysregulated MAFB expression or activity has been associated with immune and inflammatory phenotypes, kidney glomerular dysfunction, and altered cellular differentiation programs relevant to cancer biology.

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

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

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