NSD3 CRISPR Activation Plasmid (h): sc-402721-ACT

NSD3 (also known as WHSC1L1) encodes a nuclear SET domain histone lysine methyltransferase that functions as a chromatin regulator of transcriptional programs linked to cell identity and proliferation. By depositing methyl marks on histones and coordinating with other epigenetic cofactors, NSD3 contributes to nucleosome organization, enhancer activity, and RNA polymerase II–dependent gene expression. NSD3 dysregulation has been associated with altered chromatin states observed in multiple cancers, including contexts involving 8p11 amplification and aberrant transcriptional dependency. These properties make NSD3 a useful target for studying epigenetic control of oncogenic signaling, lineage programs, and chromatin-mediated gene regulation in human cells.

NSD3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NSD3 expression without altering the underlying DNA sequence.

NSD3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NSD3 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 NSD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NSD3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NSD3 locus and enabling the study of NSD3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NSD3 pathway restoration in tumor cells with silenced or reduced NSD3 expression.

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