Bex4 CRISPR Activation Plasmid (m): sc-436560-ACT

Mouse Bex4 (brain expressed X-linked 4) encodes a small, largely neuronal protein implicated in developmentally regulated transcriptional programs and neuronal differentiation. As a member of the BEX family, Bex4 is linked to signaling and stress-response networks that shape cell survival and lineage specification, with reported context-dependent interactions influencing neurotrophin-associated pathways. Expression is enriched in brain and other neuroectodermal contexts, making Bex4 a useful node for studying neuronal maturation, axon/dendrite dynamics, and activity-dependent gene regulation. Dysregulated BEX-family expression patterns have been associated with neurodevelopmental phenotypes and cancer-related transcriptional states, supporting mechanistic research into how Bex4 modulates cell-state transitions.

Bex4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Bex4 expression without altering the underlying DNA sequence.

Bex4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Bex4 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 Bex4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Bex4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Bex4 locus and enabling the study of Bex4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Bex4 pathway restoration in tumor cells with silenced or reduced Bex4 expression.

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