TRIM72 CRISPR Activation Plasmid (h): sc-403554-ACT

TRIM72 (also known as MG53) is a muscle-enriched tripartite motif (TRIM) E3 ubiquitin ligase that coordinates plasma membrane repair by sensing membrane disruption and promoting rapid vesicle recruitment to injury sites. In human cells, TRIM72 modulates protein homeostasis through ubiquitin-dependent turnover and participates in stress-response signaling linked to cytoskeletal remodeling and membrane trafficking. These processes intersect with pathways controlling myofiber integrity, calcium handling, and inflammatory signaling following cellular injury. Dysregulated TRIM72 activity has been associated with impaired membrane repair and muscle pathology, supporting its relevance in studies of myopathy, cardiometabolic stress, and tissue damage responses.

TRIM72 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRIM72 expression without altering the underlying DNA sequence.

TRIM72 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRIM72 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 TRIM72 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRIM72 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRIM72 locus and enabling the study of TRIM72-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRIM72 pathway restoration in tumor cells with silenced or reduced TRIM72 expression.

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