spectrin β III CRISPR Activation Plasmid (h): sc-402980-ACT

SPTBN2 encodes human spectrin βIII, a cytoskeletal scaffold that forms heterotetramers with α-spectrins to stabilize cortical actin networks and organize membrane microdomains. Spectrin βIII helps maintain cell shape and mechanical resilience while coordinating trafficking and localization of ion channels, receptors, and signaling complexes, linking cytoskeleton dynamics to processes such as neurite outgrowth and synaptic organization. In neurons, SPTBN2 is highly relevant to cerebellar development and Purkinje cell function, and its dysregulation is associated with neurodevelopmental and neurodegenerative phenotypes. These properties make SPTBN2 a useful node for studying cytoskeletal signaling, membrane protein compartmentalization, and stress responses that depend on actin–spectrin architecture.

spectrin β III CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPTBN2 expression without altering the underlying DNA sequence.

spectrin β III CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPTBN2 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 SPTBN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous spectrin β III expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPTBN2 locus and enabling the study of spectrin β III-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of spectrin β III pathway restoration in tumor cells with silenced or reduced SPTBN2 expression.

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