BBS2 CRISPR Activation Plasmid (h): sc-404725-ACT

BBS2 encodes a core subunit of the BBSome complex, a multimeric coat that regulates ciliary membrane protein trafficking and signaling competence of the primary cilium. Through interactions with intraflagellar transport machinery and small GTPases, BBS2 helps coordinate cargo export/import and receptor localization that influences pathways such as Hedgehog and other cilia-dependent signaling programs. Disruption of BBS2 perturbs ciliogenesis and ciliary transport, leading to altered cellular homeostasis in tissues that rely on ciliary signaling. Variants in BBS2 are implicated in Bardet–Biedl syndrome and related ciliopathy phenotypes, making it a useful target for studying cilia-mediated disease mechanisms.

BBS2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BBS2 expression without altering the underlying DNA sequence.

BBS2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BBS2 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 BBS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BBS2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BBS2 locus and enabling the study of BBS2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BBS2 pathway restoration in tumor cells with silenced or reduced BBS2 expression.

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