Bcl-3 CRISPR Activation Plasmid (h): sc-400740-ACT

BCL3 encodes Bcl-3, an atypical IκB family member that functions primarily in the nucleus as a transcriptional co-regulator of NF-κB p50/p52 homodimers. By modulating κB-dependent gene expression, Bcl-3 influences inflammatory signaling, lymphocyte activation, cell-cycle progression, and apoptosis programs, integrating cues from canonical and non-canonical NF-κB pathways. Altered BCL3 expression or regulation has been linked to immune dysregulation and oncogenic transcriptional states, including contexts where NF-κB signaling is persistently engaged. As a pathway node, Bcl-3 is frequently studied for its role in chromatin-associated transcriptional control and stimulus-dependent gene network remodeling.

Bcl-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BCL3 expression without altering the underlying DNA sequence.

Bcl-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BCL3 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 BCL3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bcl-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BCL3 locus and enabling the study of Bcl-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bcl-3 pathway restoration in tumor cells with silenced or reduced BCL3 expression.

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