MAF1 CRISPR Activation Plasmid (h): sc-418170-ACT

Human MAF1 encodes a conserved transcriptional repressor that couples nutrient and stress signaling to RNA polymerase III output, thereby regulating synthesis of tRNAs, 5S rRNA, and other small noncoding RNAs essential for protein translation and cellular growth. MAF1 activity is controlled by phosphorylation downstream of mTORC1 and related kinases, integrating metabolic status with transcriptional programs that shape proteostasis and adaptation to cellular stress. Through its influence on translational capacity, MAF1 contributes to pathways governing proliferation, energy balance, and stress responses. Dysregulation of Pol III transcription and its upstream regulators has been associated with oncogenic and metabolic phenotypes, motivating mechanistic studies of MAF1-dependent transcriptional control in disease-relevant contexts.

MAF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAF1 expression without altering the underlying DNA sequence.

MAF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAF1 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 MAF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MAF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAF1 locus and enabling the study of MAF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MAF1 pathway restoration in tumor cells with silenced or reduced MAF1 expression.

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