Desmin CRISPR Activation Plasmid (h): sc-400248-ACT

Human DES encodes desmin, a type III intermediate filament protein that forms a core structural network in skeletal, cardiac, and smooth muscle cells. Desmin filaments integrate myofibrils with the sarcolemma, mitochondria, and nucleus to support mechanical stability, force transmission, and organelle positioning during contraction and stress responses. This cytoskeletal architecture interfaces with sarcomere organization, mechanotransduction, and proteostasis pathways that maintain muscle integrity. Dysregulated DES expression or filament assembly is linked to myofibrillar myopathies and cardiomyopathies, making desmin a key target for studying muscle degeneration and cellular stress remodeling.

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

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

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