HTF9C CRISPR Activation Plasmid (h): sc-407594-ACT

Human TRMT2A encodes HTF9C, a tRNA methyltransferase implicated in post-transcriptional RNA modification through formation of 5-methyluridine at defined positions in tRNA. By shaping tRNA stability and decoding fidelity, TRMT2A supports efficient translation and proteostasis, processes that influence cell cycle progression and stress adaptation. Perturbation of tRNA modification networks has been associated with altered translational control and genome maintenance programs, making TRMT2A/HTF9C relevant to studies of proliferative signaling and cellular homeostasis. Expression and functional changes in tRNA-modifying enzymes are frequently explored in the context of cancer biology and neurobiology as contributors to dysregulated gene expression at the translational level.

HTF9C CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TRMT2A expression without altering the underlying DNA sequence.

HTF9C CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TRMT2A 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 TRMT2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HTF9C expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TRMT2A locus and enabling the study of HTF9C-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HTF9C pathway restoration in tumor cells with silenced or reduced TRMT2A expression.

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