MuRF1 CRISPR Activation Plasmid (h): sc-400797-ACT

TRIM63 encodes MuRF1, a muscle-specific RING-finger E3 ubiquitin ligase that promotes selective proteasomal turnover of sarcomeric and myofibrillar proteins during skeletal and cardiac muscle remodeling. MuRF1 integrates ubiquitin–proteasome system activity with stress-responsive signaling, including FOXO-dependent transcriptional programs and catabolic pathways that regulate protein homeostasis and contractile function. Altered TRIM63/MuRF1 expression is associated with muscle atrophy phenotypes and remodeling processes observed in chronic disease and disuse contexts, and it is frequently studied as a molecular effector of proteostasis imbalance. As a regulator of myofibril stability, MuRF1 provides a mechanistic handle for interrogating pathways linking metabolic stress, inflammation, and muscle structural adaptation.

MuRF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM63 expression without altering the underlying DNA sequence.

MuRF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM63 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 TRIM63 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MuRF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM63 locus and enabling the study of MuRF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MuRF1 pathway restoration in tumor cells with silenced or reduced TRIM63 expression.

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