Elongin A3 CRISPR Activation Plasmid (h): sc-418260-ACT

TCEB3C encodes Elongin A3, a member of the Elongin A family implicated in transcriptional control by RNA polymerase II and modulation of transcriptional elongation. Through interactions with elongin complexes and transcription-associated protein assemblies, Elongin A3 is linked to regulation of gene expression programs that coordinate cell-state transitions, stress-adaptive responses, and proteostasis-related processes. Altered expression or pathway context involving elongation control and ubiquitin-proteasome regulation is frequently explored in cancer biology and other disorders characterized by transcriptional dysregulation, making TCEB3C a useful node for mechanistic studies. Human Elongin A3 function is therefore relevant to investigating how transcriptional output is tuned across signaling pathways and chromatin states.

Elongin A3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TCEB3C expression without altering the underlying DNA sequence.

Elongin A3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TCEB3C 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 TCEB3C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Elongin A3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TCEB3C locus and enabling the study of Elongin A3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Elongin A3 pathway restoration in tumor cells with silenced or reduced TCEB3C expression.

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