Zap3 CRISPR Activation Plasmid (h): sc-412693-ACT

Human YLPM1 encodes Zap3, a nuclear protein implicated in RNA processing and transcriptional regulation through interactions with spliceosomal and chromatin-associated factors. Zap3 has been linked to control of pre-mRNA splicing, mRNA surveillance, and coordination of gene expression programs that influence cell-cycle progression and stress-responsive transcription. Perturbation of YLPM1/Zap3 expression is therefore relevant to studies of genome-wide splicing fidelity and transcriptional homeostasis in proliferative tissues. Dysregulation of these processes is frequently observed in cancer and neurodevelopmental disorders, making YLPM1 a useful node for mechanistic investigations of RNA metabolism–associated phenotypes.

Zap3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous YLPM1 expression without altering the underlying DNA sequence.

Zap3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the YLPM1 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 YLPM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Zap3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native YLPM1 locus and enabling the study of Zap3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Zap3 pathway restoration in tumor cells with silenced or reduced YLPM1 expression.

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