CBP80 CRISPR Activation Plasmid (h): sc-403736-ACT

NCBP1 encodes the human cap-binding complex subunit CBP80, a core component of the nuclear cap-binding complex that recognizes the 7-methylguanosine cap on nascent RNA polymerase II transcripts. CBP80 coordinates multiple steps of mRNA biogenesis, including pre-mRNA splicing, 3′ end processing, nuclear export, and the pioneer round of translation, and it interfaces with surveillance pathways such as nonsense-mediated mRNA decay. Through these activities, NCBP1 links transcriptional output to RNA quality control and proteome homeostasis across proliferative and differentiated cell states. Dysregulation of cap-dependent RNA processing and export is broadly relevant to oncogenic transcriptional programs and neurodevelopmental phenotypes, making CBP80 a useful node for studying RNA metabolism in disease-relevant models.

CBP80 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCBP1 expression without altering the underlying DNA sequence.

CBP80 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCBP1 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 NCBP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CBP80 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCBP1 locus and enabling the study of CBP80-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CBP80 pathway restoration in tumor cells with silenced or reduced NCBP1 expression.

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