Bcl-6 CRISPR Activation Plasmid (h2): sc-400265-ACT-2

Human BCL6 encodes the Bcl-6 transcriptional repressor, a BTB/POZ zinc-finger protein that regulates germinal center B-cell differentiation, proliferation, and survival by recruiting corepressor complexes (e.g., SMRT/NCoR, BCOR) to silence target genes. Bcl-6 integrates signals from B-cell receptor and cytokine pathways to modulate programs controlling DNA damage responses, apoptosis, and cell-cycle checkpoints, thereby shaping antibody affinity maturation. Dysregulated BCL6 expression or activity is strongly associated with germinal center–derived lymphoid malignancies and altered immune regulation, making it a key node in studies of oncogenic transcriptional networks. Gene editing of BCL6 is widely used to dissect transcriptional circuitry, map enhancer–promoter dependencies, and evaluate functional consequences of regulatory variants in immune and cancer model systems.

Bcl-6 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous BCL6 expression without altering the underlying DNA sequence.

Bcl-6 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BCL6 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 BCL6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bcl-6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BCL6 locus and enabling the study of Bcl-6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bcl-6 pathway restoration in tumor cells with silenced or reduced BCL6 expression.

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