MafA CRISPR Activation Plasmid (h): sc-401480-ACT

Human MAFA encodes MafA, a basic leucine zipper transcription factor that binds Maf recognition elements to regulate glucose-stimulated insulin secretion programs and pancreatic β-cell maturation. MafA integrates nutrient and stress-responsive signaling to control transcriptional networks involved in insulin gene expression, secretory granule function, and β-cell identity maintenance. Dysregulated MAFA activity has been linked to impaired β-cell function and altered endocrine differentiation, making it relevant for studies of diabetes-associated transcriptional circuitry and islet cell biology. As a lineage-defining regulator, MafA is also used to interrogate gene regulatory networks governing endocrine cell fate decisions and metabolic homeostasis.

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

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

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