IFT172 CRISPR Activation Plasmid (h): sc-404379-ACT

IFT172 encodes an intraflagellar transport (IFT) complex B component that is essential for anterograde trafficking within primary cilia, supporting axoneme assembly, ciliary maintenance, and signal transduction. By enabling ciliary cargo movement, IFT172 influences cilia-dependent developmental and homeostatic pathways, including Hedgehog signaling and other receptor-mediated processes that rely on intact ciliary architecture. Disruption or dysregulation of IFT172 is associated with ciliopathy-related phenotypes, linking altered ciliary transport to defects in tissue patterning, sensory function, and organ development. As a ciliary transport factor, IFT172 is widely used to interrogate mechanisms of ciliogenesis, cilia-mediated signaling dynamics, and genotype–phenotype relationships in human cell models.

IFT172 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous IFT172 expression without altering the underlying DNA sequence.

IFT172 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the IFT172 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 IFT172 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous IFT172 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native IFT172 locus and enabling the study of IFT172-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of IFT172 pathway restoration in tumor cells with silenced or reduced IFT172 expression.

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