MLL CRISPR Activation Plasmid (h): sc-401307-ACT

KMT2A (MLL) encodes a SET-domain histone lysine methyltransferase that catalyzes H3K4 methylation to regulate chromatin accessibility and transcriptional programs controlling hematopoietic development and cell-fate specification. MLL functions in large nuclear complexes that coordinate epigenetic writing with transcriptional elongation at promoters and enhancers, impacting pathways such as HOX gene regulation, DNA damage responses, and differentiation. Dysregulation of KMT2A through altered expression or genomic rearrangements is strongly linked to aberrant transcriptional states in leukemia and other malignancies, making MLL a central node for studying epigenetic control of oncogenic gene expression. Its broad role in chromatin-mediated regulation also supports investigation of context-dependent effects on proliferation, lineage commitment, and transcriptional network stability in human cells.

MLL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KMT2A expression without altering the underlying DNA sequence.

MLL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KMT2A 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 KMT2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MLL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KMT2A locus and enabling the study of MLL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MLL pathway restoration in tumor cells with silenced or reduced KMT2A expression.

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