Msi1 CRISPR Activation Plasmid (h): sc-404014-ACT

MSI1 encodes Musashi RNA-binding protein 1 (Msi1), a conserved post-transcriptional regulator that binds sequence motifs in target mRNAs to modulate translation, stability, and cell fate decisions. In human cells, Msi1 supports stem/progenitor maintenance and influences differentiation programs by shaping signaling outputs in pathways such as Notch and Wnt/β-catenin, as well as broader RNA processing networks that control proliferation and lineage commitment. Aberrant MSI1 expression has been linked to dysregulated growth and impaired differentiation states in multiple disease contexts, making it a useful marker and mechanistic entry point for studying oncogenic-like transcriptional and translational circuitry. As an RNA-binding protein, Msi1 is also relevant to investigations of stress responses and RNA granule-associated regulation that can rewire gene expression without altering DNA sequence.

Msi1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MSI1 expression without altering the underlying DNA sequence.

Msi1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MSI1 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 MSI1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Msi1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MSI1 locus and enabling the study of Msi1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Msi1 pathway restoration in tumor cells with silenced or reduced MSI1 expression.

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