SH2D1A CRISPR Activation Plasmid (h): sc-404747-ACT

SH2D1A encodes the SLAM-associated adaptor protein (SAP), an SH2 domain–containing signaling adaptor that couples SLAM family receptors to downstream tyrosine kinase pathways in lymphocytes. By mediating receptor-proximal signaling events, SAP regulates cytotoxic lymphocyte activation, T cell–B cell interactions, and germinal center dynamics that shape adaptive immune responses. SH2D1A activity influences immune synapse formation and effector functions through modulation of phosphorylation cascades and checkpoint-like receptor signaling. Pathogenic variation in SH2D1A is linked to primary immunodeficiency phenotypes, including X-linked lymphoproliferative disease, highlighting its importance in immune homeostasis and antiviral immunity research.

SH2D1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SH2D1A expression without altering the underlying DNA sequence.

SH2D1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SH2D1A 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 SH2D1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SH2D1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SH2D1A locus and enabling the study of SH2D1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SH2D1A pathway restoration in tumor cells with silenced or reduced SH2D1A expression.

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