CBP20 CRISPR Activation Plasmid (h): sc-404124-ACT

Human NCBP2 encodes CBP20, the small subunit of the nuclear cap-binding complex that recognizes the 7-methylguanosine cap on nascent RNA polymerase II transcripts. Through association with CBP80 and interaction with splicing, mRNA export, and RNA surveillance factors, CBP20 helps coordinate pre-mRNA processing, nuclear export competence, and quality control pathways such as nonsense-mediated decay. CBP20-dependent cap recognition influences gene expression programs during cell cycle progression and stress responses by shaping transcript maturation and stability. Dysregulation of cap-dependent RNA processing and export is frequently linked to altered proliferation and genome maintenance phenotypes, making NCBP2 a relevant target for mechanistic studies of RNA metabolism in disease-associated contexts.

CBP20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCBP2 expression without altering the underlying DNA sequence.

CBP20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCBP2 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 NCBP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CBP20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCBP2 locus and enabling the study of CBP20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CBP20 pathway restoration in tumor cells with silenced or reduced NCBP2 expression.

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