coilin CRISPR Activation Plasmid (h): sc-400927-ACT

Human COIL encodes coilin, a signature scaffolding protein of Cajal bodies that coordinates the assembly and maturation of small nuclear and small nucleolar ribonucleoproteins, supporting pre-mRNA splicing and RNA processing. Coilin interacts with spliceosomal components and guides the trafficking of RNA-protein complexes between nuclear subdomains, linking nuclear architecture to gene expression regulation. Through these roles, COIL contributes to pathways governing RNA metabolism, ribosome biogenesis, and cell cycle–coupled nuclear organization. Altered coilin localization or Cajal body integrity has been observed in contexts of cellular stress and malignant transformation, making COIL a useful target for mechanistic studies of nuclear bodies and transcriptome regulation.

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

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

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