SMG1 CRISPR Activation Plasmid (h): sc-404760-ACT

SMG1 encodes a PI3K-related serine/threonine kinase that functions as a core component of the nonsense-mediated mRNA decay (NMD) pathway by phosphorylating UPF1 and coordinating surveillance of premature termination codon–containing transcripts. Through regulation of mRNA stability and translation, SMG1 contributes to proteostasis, cellular stress responses, and genome integrity programs that intersect with DNA damage signaling and cell cycle control. Perturbation of SMG1-dependent NMD can reshape transcriptome quality control, influencing differentiation states and stress-adaptive gene expression networks. Dysregulated SMG1 activity has been linked to altered RNA surveillance and signaling phenotypes observed in multiple disease contexts, supporting its use as a mechanistic node for studying RNA metabolism and stress-related pathways.

SMG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMG1 expression without altering the underlying DNA sequence.

SMG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMG1 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 SMG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMG1 locus and enabling the study of SMG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMG1 pathway restoration in tumor cells with silenced or reduced SMG1 expression.

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