Ly6G6d CRISPR Activation Plasmid (h): sc-403030-ACT

LY6G6D encodes Ly6G6d, a glycosylphosphatidylinositol (GPI)-anchored member of the Ly6/uPAR family enriched in immune-related contexts and thought to influence cell–cell interactions at the plasma membrane. As a surface-associated protein, Ly6G6d is studied for roles in shaping signal transduction, membrane microdomain organization, and immune cell communication within inflammatory and tumor-associated microenvironments. Altered LY6G6D expression has been reported across multiple cancer and immune-oncology datasets, supporting its use as a molecular marker in studies of immune infiltration and epithelial–immune crosstalk. These features make LY6G6D a relevant target for dissecting pathways that link surface antigen regulation to immune signaling and disease-associated transcriptional programs.

Ly6G6d CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LY6G6D expression without altering the underlying DNA sequence.

Ly6G6d CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LY6G6D 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 LY6G6D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ly6G6d expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LY6G6D locus and enabling the study of Ly6G6d-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ly6G6d pathway restoration in tumor cells with silenced or reduced LY6G6D expression.

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