BID CRISPR Activation Plasmid (h): sc-400510-ACT

Human BID encodes a BH3-only BCL-2 family protein that links extrinsic death receptor signaling to the intrinsic mitochondrial apoptosis pathway. Following caspase-8 cleavage to truncated BID (tBID), it promotes BAX/BAK-dependent mitochondrial outer membrane permeabilization, cytochrome c release, and downstream caspase activation, thereby integrating apoptotic inputs with cellular stress responses. BID activity intersects with p53 signaling, NF-κB-regulated survival pathways, and mitochondrial dynamics that shape cell fate decisions. Dysregulated BID expression or processing has been associated with altered apoptotic thresholds observed in cancer biology, immune homeostasis, and tissue injury models, making it a useful node for mechanistic studies of cell death regulation.

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

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

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