Tafazzin CRISPR Activation Plasmid (h): sc-403588-ACT

Human TAZ encodes tafazzin, a mitochondrial phospholipid transacylase that remodels cardiolipin to maintain inner mitochondrial membrane architecture and optimize oxidative phosphorylation. By regulating cardiolipin acyl-chain composition, tafazzin supports respiratory chain supercomplex stability, mitochondrial dynamics, and mitophagy-associated quality control. Perturbation of TAZ-mediated lipid remodeling is linked to mitochondrial dysfunction and altered energy metabolism, with strong relevance to Barth syndrome and related cardiomyopathy and myopathy phenotypes. TAZ is therefore widely studied in pathways connecting mitochondrial bioenergetics, redox homeostasis, and membrane lipid signaling.

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

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

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