KIAA0100 CRISPR Activation Plasmid (h): sc-408006-ACT

Human KIAA0100 (also known as BCOX1) encodes a large, poorly characterized cytoplasmic protein implicated in the regulation of cell growth and survival programs. Transcriptomic and functional studies have linked KIAA0100 to processes such as proliferation, stress adaptation, and cytoskeletal or vesicular organization, suggesting roles in coordinated signaling networks that support cellular fitness. Altered KIAA0100 expression has been reported across multiple tumor types and is associated with aggressive phenotypes, making it a useful molecular handle for studying oncogenic transcriptional states. Due to limited mechanistic annotation, KIAA0100 is frequently investigated using perturbation approaches to map downstream pathways and interaction partners.

KIAA0100 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KIAA0100 expression without altering the underlying DNA sequence.

KIAA0100 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KIAA0100 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 KIAA0100 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIAA0100 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KIAA0100 locus and enabling the study of KIAA0100-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIAA0100 pathway restoration in tumor cells with silenced or reduced KIAA0100 expression.

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