c-Maf CRISPR Activation Plasmid (m2): sc-421531-ACT-2

The mouse Maf gene encodes the transcription factor c-Maf, a basic leucine zipper (bZIP) regulator that binds MAF recognition elements to control context-dependent gene expression programs. c-Maf integrates signals from immune and developmental pathways to modulate cellular differentiation and function, including T cell polarization (notably Th2 and Tfh programs), macrophage activation states, and lineage-specific transcriptional networks in tissues such as lens and pancreatic islets. Dysregulated c-Maf activity has been linked to altered immune homeostasis and inflammatory phenotypes and is frequently studied in models of lymphoid transformation and metabolic or ocular dysfunction. Gene editing of Maf in murine cells or mice enables mechanistic dissection of enhancer usage, transcriptional circuitry, and downstream cytokine and lineage markers using CRISPR-based knockout, knock-in, or regulatory perturbation approaches coupled to RNA-seq, ChIP-seq, and single-cell profiling.

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

c-Maf CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Maf 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 Maf transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous c-Maf expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Maf locus and enabling the study of c-Maf-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of c-Maf pathway restoration in tumor cells with silenced or reduced Maf expression.

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