plexin-B1 CRISPR Activation Plasmid (h): sc-401840-ACT

PLXNB1 encodes plexin-B1, a transmembrane receptor for class 4 semaphorins that coordinates contact-dependent signaling to regulate cytoskeletal remodeling, directional cell migration, and axon guidance. Plexin-B1 integrates cues through Rho family GTPases and associated effectors to influence actin dynamics, adhesion turnover, and cell polarity programs, with downstream impacts on tissue patterning and barrier organization. In human biology, altered PLXNB1 expression or signaling has been linked to dysregulated motility and invasive behavior in cancer models, and to neurodevelopmental processes where semaphorin–plexin pathways shape neuronal connectivity. These features make plexin-B1 a useful node for studying semaphorin signaling, Rho/ROCK-linked remodeling, and context-specific changes in cell–cell and cell–matrix interactions.

plexin-B1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLXNB1 expression without altering the underlying DNA sequence.

plexin-B1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLXNB1 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 PLXNB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous plexin-B1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLXNB1 locus and enabling the study of plexin-B1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of plexin-B1 pathway restoration in tumor cells with silenced or reduced PLXNB1 expression.

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