VPRBP CRISPR Activation Plasmid (h): sc-403283-ACT

Human DCAF1, also known as VPRBP, encodes a substrate receptor for the CUL4–DDB1 E3 ubiquitin ligase complex that directs ubiquitination and proteasomal turnover of diverse nuclear proteins. Through this adaptor function, VPRBP helps coordinate DNA replication licensing, chromatin regulation, DNA damage responses, and cell-cycle progression, linking ubiquitin-mediated proteostasis to transcriptional control. VPRBP has also been implicated in signaling crosstalk with kinases and in immune and stress-response programs via regulated degradation of pathway components. Dysregulation of DCAF1/VPRBP-dependent ubiquitination is associated with altered proliferation and genome maintenance, making it a useful node for studying oncogenic signaling, chromatin-dependent gene regulation, and host–pathogen interactions.

VPRBP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DCAF1 expression without altering the underlying DNA sequence.

VPRBP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DCAF1 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 DCAF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VPRBP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DCAF1 locus and enabling the study of VPRBP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VPRBP pathway restoration in tumor cells with silenced or reduced DCAF1 expression.

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