MafB CRISPR Activation Plasmid (m2): sc-421335-ACT-2

Mouse Mafb encodes the MafB transcription factor, a basic leucine zipper protein that regulates lineage specification and terminal differentiation in macrophages/monocytes, pancreatic endocrine cells, and developing kidney podocytes through sequence-specific control of gene expression. MafB functions within transcriptional and epigenetic programs downstream of developmental and immune signaling pathways, influencing macrophage polarization, cytokine-responsive networks, and maintenance of podocyte structure and filtration barrier integrity. Dysregulated Mafb activity has been linked to altered immune homeostasis and defects in organogenesis, and MafB expression changes are frequently examined in models of glomerular injury, diabetes-associated islet dysfunction, and inflammatory stress. Gene editing of Mafb in mouse supports mechanistic studies of transcriptional circuitry, cell fate decisions, and tissue-specific pathology using in vivo models, primary cells, and differentiated stem cell systems.

MafB CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Mafb expression without altering the underlying DNA sequence.

MafB CRISPR Activation Plasmid (m2) 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.