SART-3 CRISPR Activation Plasmid (h): sc-411332-ACT

Human SART3 encodes SART-3, a nuclear RNA-binding protein implicated in spliceosome assembly and recycling, supporting U4/U6 snRNP dynamics and pre-mRNA splicing fidelity. Through its roles in RNA processing and nuclear organization, SART-3 contributes to regulation of gene expression programs that influence cell-cycle progression, stress responses, and proteostasis. Altered splicing factor activity and spliceosomal dysregulation are recurrent features of tumor biology and immune-related phenotypes, making SART3 a useful node for studying transcriptome remodeling in disease-relevant contexts. As an immunogenic antigen described in multiple malignancies, SART-3 has also been used to explore links between RNA processing and tumor-associated antigen presentation without implying clinical utility.

SART-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SART3 expression without altering the underlying DNA sequence.

SART-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SART3 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 SART3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SART-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SART3 locus and enabling the study of SART-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SART-3 pathway restoration in tumor cells with silenced or reduced SART3 expression.

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