ASH1L CRISPR Activation Plasmid (h): sc-402901-ACT

ASH1L (absent, small, or homeotic discs 1-like) encodes a nuclear histone lysine methyltransferase that deposits H3K36 methylation to support transcriptional elongation and maintain cell type–specific gene expression programs. Through its chromatin-reader and catalytic domains, ASH1L helps balance Polycomb-mediated repression and activation at developmental loci, influencing processes such as neuronal differentiation and lineage commitment. ASH1L-regulated epigenetic control intersects with pathways governing chromatin remodeling, enhancer activity, and transcriptional fidelity. Genetic variation or dysregulation of ASH1L has been associated with neurodevelopmental phenotypes and altered gene regulatory networks, making it a relevant target for mechanistic studies of chromatin-driven transcription.

ASH1L CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ASH1L expression without altering the underlying DNA sequence.

ASH1L CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ASH1L 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 ASH1L transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASH1L expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ASH1L locus and enabling the study of ASH1L-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASH1L pathway restoration in tumor cells with silenced or reduced ASH1L expression.

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