MLL2 CRISPR Activation Plasmid (h): sc-401659-ACT

KMT2D (MLL2) encodes a SET domain–containing histone lysine methyltransferase that catalyzes H3K4 mono- and di-methylation at enhancers, shaping chromatin accessibility and lineage-specific transcriptional programs. Through interactions with COMPASS-like complexes, MLL2 coordinates enhancer activation, RNA polymerase II recruitment, and long-range gene regulation important for development and cell identity. Disruption of KMT2D-associated epigenetic control is linked to altered differentiation states, DNA damage responses, and broad transcriptional dysregulation observed across multiple cancer and developmental disease contexts. As a regulator of enhancer landscapes, KMT2D is widely studied in pathways governing cell fate specification, immune signaling, and chromatin-dependent control of proliferation.

MLL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KMT2D expression without altering the underlying DNA sequence.

MLL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KMT2D 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 KMT2D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MLL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KMT2D locus and enabling the study of MLL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MLL2 pathway restoration in tumor cells with silenced or reduced KMT2D expression.

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