MLL5 CRISPR Activation Plasmid (h): sc-403300-ACT

KMT2E encodes the histone lysine methyltransferase MLL5, a chromatin-associated regulator that influences transcriptional programs controlling cell cycle progression, hematopoietic differentiation, and lineage commitment. Although its catalytic activity is debated relative to other KMT2 family members, MLL5 participates in epigenetic regulation through interactions with chromatin and transcriptional complexes, impacting histone modification landscapes and gene expression stability. Dysregulation of KMT2E/MLL5 has been linked to altered proliferation and differentiation states and is frequently studied in the context of myeloid biology, genome-wide chromatin control, and cancer-associated transcriptional rewiring. These properties make KMT2E a useful node for dissecting epigenetic pathways, transcriptional circuitry, and phenotype-to-expression relationships in human cells.

MLL5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KMT2E expression without altering the underlying DNA sequence.

MLL5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KMT2E 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 KMT2E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MLL5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KMT2E locus and enabling the study of MLL5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MLL5 pathway restoration in tumor cells with silenced or reduced KMT2E expression.

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