emerin CRISPR Activation Plasmid (h): sc-401048-ACT

Human EMD encodes emerin, a LEM-domain inner nuclear membrane protein that anchors the nuclear lamina and couples the nucleus to cytoskeletal forces through interactions with lamins, BAF, and LINC-associated components. Emerin contributes to nuclear envelope organization, mechanotransduction, and chromatin tethering, influencing transcriptional regulation, DNA damage responses, and cell cycle progression. Perturbation of emerin-dependent nuclear architecture is linked to tissue-specific pathology in striated muscle and cardiac conduction systems, making EMD a widely used model for nuclear envelope-associated disease biology. As a scaffold at the nuclear periphery, emerin is frequently studied in pathways governing nuclear stiffness, genome stability, and differentiation-dependent gene expression programs.

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

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

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