LSm1 CRISPR Activation Plasmid (h): sc-403980-ACT

Human LSM1 encodes LSm1, an Sm-like RNA-binding protein that assembles in the cytoplasmic LSM1–7 complex to regulate mRNA turnover. LSm1 promotes 5′-to-3′ mRNA decay by supporting decapping and coordinating with P-bodies, thereby shaping transcript stability and gene expression programs linked to proliferation and stress responses. Through its role in post-transcriptional control, altered LSM1 activity can influence pathways connected to oncogenic transformation and dysregulated RNA metabolism, making it relevant for mechanistic studies of disease-associated gene expression states. LSm1 is also used as a marker and functional node for investigating RNA processing defects and their downstream impacts on cell behavior.

LSm1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LSM1 expression without altering the underlying DNA sequence.

LSm1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LSM1 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 LSM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LSm1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LSM1 locus and enabling the study of LSm1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LSm1 pathway restoration in tumor cells with silenced or reduced LSM1 expression.

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