MSG1 CRISPR Activation Plasmid (h): sc-404984-ACT

Human CITED1 (MSG1) encodes a nuclear transcriptional co-regulator that binds CBP/p300 and modulates signal-dependent gene expression programs. It functions in hormone and growth factor–responsive transcription, influencing differentiation and proliferation through pathways such as TGF-β/SMAD and estrogen receptor–associated networks. CITED1 is highly relevant in developmental biology and epithelial lineage specification, where it can tune chromatin-dependent transcriptional outputs. Dysregulated CITED1 expression has been reported across multiple tumor contexts and is studied as a marker and mechanistic node in oncogenic transcriptional reprogramming.

MSG1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CITED1 expression without altering the underlying DNA sequence.

MSG1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CITED1 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 CITED1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MSG1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CITED1 locus and enabling the study of MSG1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MSG1 pathway restoration in tumor cells with silenced or reduced CITED1 expression.

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