Nanos2 CRISPR Activation Plasmid (h): sc-404123-ACT

Human NANOS2 encodes Nanos2, a conserved RNA-binding protein best known for regulating germ cell fate by controlling post-transcriptional gene expression programs. Nanos2 participates in mRNA stability and translational repression networks that help maintain germline identity and restrain inappropriate differentiation, integrating with pathways that govern cell-cycle progression, meiosis entry, and developmental timing. Altered NANOS family activity has been associated with defects in germ cell development and dysregulated reproductive biology, and NANOS2 expression is often examined as a marker of lineage specification in developmental and stem cell models. These features make NANOS2 a useful node for studying RNA regulons, developmental gene networks, and context-dependent transcriptional states.

Nanos2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NANOS2 expression without altering the underlying DNA sequence.

Nanos2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NANOS2 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 NANOS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nanos2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NANOS2 locus and enabling the study of Nanos2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nanos2 pathway restoration in tumor cells with silenced or reduced NANOS2 expression.

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