LSD1 CRISPR Activation Plasmid (m): sc-430289-ACT

Mouse Kdm1a encodes lysine-specific demethylase 1 (LSD1/KDM1A), a FAD-dependent histone demethylase that primarily removes H3K4me1/2 and H3K9me1/2 marks to modulate chromatin accessibility and transcriptional programs. LSD1 functions within multiprotein repressor and activator complexes, including CoREST and NuRD, linking epigenetic remodeling to RNA polymerase II–dependent gene regulation during development, lineage commitment, and cell-cycle control. By shaping enhancer and promoter states, Kdm1a influences pathways governing differentiation, metabolism, and stress responses in multiple tissues. Dysregulated LSD1 activity is widely used as a mechanistic entry point for studying aberrant epigenetic regulation associated with oncogenic transcriptional states and neurodevelopmental phenotypes in experimental models.

LSD1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kdm1a expression without altering the underlying DNA sequence.

LSD1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kdm1a 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 Kdm1a transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LSD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kdm1a locus and enabling the study of LSD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LSD1 pathway restoration in tumor cells with silenced or reduced Kdm1a expression.

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