Siglec-1 CRISPR Activation Plasmid (h): sc-402497-ACT

SIGLEC1 encodes Siglec-1 (CD169), a sialic acid–binding immunoglobulin-like lectin selectively expressed on macrophages and specific dendritic cell populations that mediates recognition and capture of sialylated glycoconjugates on host cells and pathogens. Through its role in adhesion, endocytosis, and antigen handling, Siglec-1 contributes to innate immune sensing, cell–cell interactions, and shaping inflammatory responses within tissue microenvironments. It is commonly linked to type I interferon–driven activation programs and macrophage polarization states, making it a useful marker and functional node in myeloid immune biology. Dysregulated SIGLEC1 expression has been associated with chronic inflammatory and autoimmune conditions, infectious disease processes, and tumor-associated macrophage phenotypes, supporting its relevance in immunology and disease-mechanism research.

Siglec-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SIGLEC1 expression without altering the underlying DNA sequence.

Siglec-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SIGLEC1 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 SIGLEC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Siglec-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SIGLEC1 locus and enabling the study of Siglec-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Siglec-1 pathway restoration in tumor cells with silenced or reduced SIGLEC1 expression.

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