Nanos1 CRISPR Activation Plasmid (h): sc-404414-ACT

Human NANOS1 encodes Nanos1, an evolutionarily conserved RNA-binding protein that regulates post-transcriptional gene expression in germ cells by modulating mRNA stability and translation. NANOS1 participates in germline maintenance and differentiation programs through repression of specific transcripts, supporting cell fate decisions, survival, and cell-cycle control. Dysregulated NANOS1 activity has been associated with impaired spermatogenesis and male infertility phenotypes, and altered expression has been reported in select tumor contexts where germline-associated pathways are ectopically engaged. These properties make NANOS1 a useful node for studying RNA regulatory networks, developmental gene control, and mechanisms that link germline factors to cellular plasticity.

Nanos1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NANOS1 expression without altering the underlying DNA sequence.

Nanos1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NANOS1 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 NANOS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nanos1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NANOS1 locus and enabling the study of Nanos1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nanos1 pathway restoration in tumor cells with silenced or reduced NANOS1 expression.

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