C9orf41 CRISPR Activation Plasmid (h): sc-409368-ACT

CARNMT1 encodes the human C9orf41 protein, an S-adenosyl-L-methionine–dependent methyltransferase implicated in RNA metabolism through methylation of the cap-adjacent adenosine (Am) to form N6,2′-O-dimethyladenosine (m6Am) at the 5′ end of select transcripts. This epitranscriptomic modification can influence mRNA stability and translation efficiency, linking C9orf41 activity to post-transcriptional gene regulation and proteostasis. Altered RNA cap-associated methylation and dysregulated RNA processing pathways are recurrent features across diverse cancer and neurological disease datasets, making CARNMT1 a useful node for mechanistic studies of transcription–translation coupling. Modulating C9orf41 expression supports interrogation of how m6Am dynamics shape cell-state transitions, stress responses, and pathway-specific gene expression programs.

C9orf41 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CARNMT1 expression without altering the underlying DNA sequence.

C9orf41 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CARNMT1 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 CARNMT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous C9orf41 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CARNMT1 locus and enabling the study of C9orf41-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of C9orf41 pathway restoration in tumor cells with silenced or reduced CARNMT1 expression.

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