ZC3H13 CRISPR Activation Plasmid (m): sc-426493-ACT

Zc3h13 encodes ZC3H13, a CCCH-type zinc finger RNA-binding protein that functions as a core component of the mRNA N6-methyladenosine (m6A) writer-associated machinery. In mouse cells, ZC3H13 supports nuclear localization and scaffolding of m6A regulatory complexes, influencing co-transcriptional RNA processing, alternative splicing, mRNA stability, and gene expression programs linked to cell fate decisions. Through these roles, ZC3H13 intersects with pathways governing transcriptional regulation and post-transcriptional control that shape development and tissue homeostasis. Dysregulation of m6A pathway components, including ZC3H13-associated networks, has been implicated in altered differentiation states and disease-relevant transcriptional signatures, making Zc3h13 a useful target for mechanistic studies.

ZC3H13 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Zc3h13 expression without altering the underlying DNA sequence.

ZC3H13 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Zc3h13 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 Zc3h13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ZC3H13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Zc3h13 locus and enabling the study of ZC3H13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ZC3H13 pathway restoration in tumor cells with silenced or reduced Zc3h13 expression.

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