BTBD14B CRISPR Activation Plasmid (h): sc-410250-ACT

NACC1 encodes a nuclear BTB/POZ domain–containing transcriptional regulator implicated in chromatin-associated control of gene expression programs that influence proliferation, differentiation, and cellular stress responses. Through interactions with corepressor complexes and transcriptional machinery, NACC1 has been linked to pathways governing cell-cycle progression, epithelial–mesenchymal transition, and metabolic adaptation. Altered NACC1 activity has been reported in multiple disease contexts, particularly in cancer biology, where it is associated with changes in tumor cell fitness, invasion-related phenotypes, and therapy response signatures. These features make NACC1/BTBD14B a useful node for dissecting transcriptional network rewiring and lineage-state regulation in human model systems.

BTBD14B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NACC1 expression without altering the underlying DNA sequence.

BTBD14B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NACC1 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 NACC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BTBD14B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NACC1 locus and enabling the study of BTBD14B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BTBD14B pathway restoration in tumor cells with silenced or reduced NACC1 expression.

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