TERT CRISPR Activation Plasmid (h): sc-400316-ACT

Human TERT encodes telomerase reverse transcriptase, the catalytic subunit of telomerase that extends telomeric repeats to maintain chromosome end integrity and enable long-term replicative capacity. TERT activity coordinates with the telomere maintenance network, including shelterin complex regulation, DNA replication stress responses, and DNA damage signaling pathways such as ATM/ATR that are engaged when telomeres become dysfunctional. Dysregulated telomerase expression contributes to altered cellular senescence, genome instability, and immortalization phenotypes, while inherited or acquired perturbations in telomere biology are linked to diverse disorders characterized by premature telomere shortening. As a central node in telomere homeostasis, TERT is widely studied in models of stem cell self-renewal, replicative aging, and telomere-driven genome maintenance.

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

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

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