SP140 CRISPR Activation Plasmid (h): sc-409816-ACT

SP140 encodes a nuclear speckled protein containing PHD and bromodomains that functions as a chromatin reader and transcriptional regulator, with enriched expression in immune lineages. It participates in epigenetic control of gene programs linked to innate and adaptive immune responses, influencing pathways such as interferon signaling and inflammatory transcriptional networks. Through interactions with modified histones and chromatin-associated complexes, SP140 helps shape lineage-specific gene expression and cellular activation states. Genetic and expression studies have associated SP140 dysregulation with immune-mediated and inflammatory disease susceptibility, supporting its relevance for mechanistic research in immunology and chromatin biology.

SP140 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SP140 expression without altering the underlying DNA sequence.

SP140 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SP140 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 SP140 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SP140 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SP140 locus and enabling the study of SP140-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SP140 pathway restoration in tumor cells with silenced or reduced SP140 expression.

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