C19orf66 CRISPR Activation Plasmid (h): sc-408037-ACT

C19orf66 encodes an interferon-stimulated protein (also known as shiftless, SHFL) that restricts the replication of diverse viruses by modulating viral RNA translation and altering the accumulation of viral proteins. It participates in innate immune signaling downstream of type I interferon and contributes to antiviral effector programs that reshape RNA metabolism and protein synthesis in infected cells. C19orf66 has been linked to pathways controlling RNA stability and translational control, and its activity intersects with cellular stress responses that influence host–pathogen interactions. Dysregulation of interferon-stimulated gene networks, including C19orf66, is relevant to studies of chronic inflammation, viral susceptibility, and immune-associated disease phenotypes.

C19orf66 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C19orf66 expression without altering the underlying DNA sequence.

C19orf66 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C19orf66 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 C19orf66 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C19orf66 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C19orf66 locus and enabling the study of C19orf66-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C19orf66 pathway restoration in tumor cells with silenced or reduced C19orf66 expression.

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