N-RAP CRISPR Activation Plasmid (h): sc-408143-ACT

Human NRAP encodes N-RAP, a striated muscle–enriched cytoskeletal scaffold that coordinates myofibril assembly by linking actin filaments with α-actinin–containing structures at developing Z-discs and costameres. Through interactions with sarcomeric and membrane-associated proteins, N-RAP supports organization of contractile architecture and mechanotransduction processes that shape muscle cell differentiation and stability. Altered NRAP expression or variants have been associated with cardiac and skeletal muscle dysfunction in genetic and remodeling contexts, making it relevant to pathways governing sarcomere maintenance, stress adaptation, and muscle-specific gene programs. NRAP is therefore frequently studied in models of myopathy and cardiomyopathy, as well as in systems probing cytoskeletal remodeling and muscle maturation.

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

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

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