SIRT6 CRISPR Activation Plasmid (m): sc-424467-ACT

Mouse Sirt6 encodes SIRT6, a nuclear NAD+-dependent deacylase that regulates chromatin structure and transcription through histone H3 deacetylation and ADP-ribosylation activities. SIRT6 integrates metabolic sensing with genome maintenance by modulating DNA double-strand break repair, telomere stability, and replication stress responses, and it influences glucose and lipid metabolism via control of glycolytic and inflammatory gene programs. Functionally, SIRT6 intersects with pathways involving NF-κB signaling, oxidative stress responses, and chromatin remodeling, shaping cellular aging and stress resilience. Dysregulated SIRT6 activity has been linked in research to age-associated phenotypes, metabolic imbalance, and inflammatory states, making it a key node for mechanistic studies in mouse systems.

SIRT6 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Sirt6 expression without altering the underlying DNA sequence.

SIRT6 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Sirt6 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 Sirt6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SIRT6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Sirt6 locus and enabling the study of SIRT6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SIRT6 pathway restoration in tumor cells with silenced or reduced Sirt6 expression.

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