LSM8 CRISPR Activation Plasmid (h): sc-404816-ACT

Human LSM8 encodes an Sm-like RNA-binding protein that is a core component of the U6 small nuclear ribonucleoprotein and the LSm2–8 ring, supporting U6 snRNA stability, snRNP biogenesis, and spliceosome assembly during pre-mRNA splicing. Through these functions, LSM8 contributes to accurate intron removal and transcriptome integrity, linking it to RNA processing networks that shape cell-cycle progression and stress-responsive gene expression programs. Perturbation of spliceosomal and snRNP homeostasis is broadly associated with aberrant mRNA isoforms and dysregulated gene expression in multiple disease contexts, making LSM8 a useful node for mechanistic studies of splicing-dependent phenotypes. Its essential role in RNA metabolism also provides a defined entry point to interrogate how core splicing factors influence cellular state and proteome diversity.

LSM8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LSM8 expression without altering the underlying DNA sequence.

LSM8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LSM8 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 LSM8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LSM8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LSM8 locus and enabling the study of LSM8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LSM8 pathway restoration in tumor cells with silenced or reduced LSM8 expression.

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