SLBP CRISPR Activation Plasmid (h): sc-403637-ACT

Stem-loop binding protein (SLBP) is a conserved RNA-binding factor that recognizes the 3′ stem–loop structure of replication-dependent histone mRNAs, coordinating their processing, nuclear export, translation, and rapid degradation at the end of S phase. By coupling histone supply to DNA replication, SLBP supports chromatin assembly, genome stability, and orderly cell-cycle progression, integrating with S-phase checkpoints and DNA damage response pathways. Dysregulated SLBP activity can perturb histone homeostasis, leading to replication stress, altered chromatin landscapes, and transcriptional instability. As a result, SLBP is frequently examined in contexts of proliferative biology and tumor-associated cell-cycle deregulation as a mechanistic node linking RNA processing to chromatin maintenance.

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

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

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