Acrosin CRISPR Activation Plasmid (h): sc-402292-ACT

Human ACR encodes acrosin, a trypsin-like serine protease stored as proacrosin within the acrosome of mature spermatozoa. Upon acrosome reaction, acrosin participates in zona pellucida binding and localized proteolysis that supports sperm penetration of the oocyte investments, integrating with fertilization-associated processes such as acrosomal exocytosis, protease activation cascades, and extracellular matrix remodeling. Dysregulated acrosin activity or impaired acrosome function has been associated with male-factor infertility phenotypes, including reduced fertilization capacity and abnormal sperm–zona interactions. As a germ cell–enriched protease, ACR is also used as a marker for studying spermatogenesis, acrosome biogenesis, and sperm functional maturation.

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

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

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