DCBLD1 CRISPR Activation Plasmid (h): sc-404343-ACT

DCBLD1 (discoidin, CUB and LCCL domain containing 1) encodes a single-pass transmembrane protein implicated in cell–cell and cell–matrix communication through extracellular domain-mediated interactions. Human DCBLD1 has been linked to regulation of signaling networks that influence adhesion, motility, and receptor tyrosine kinase-associated pathways, supporting roles in cytoskeletal remodeling and context-dependent transcriptional programs. Altered DCBLD1 expression has been reported in datasets spanning multiple tumor types and vascular-associated phenotypes, suggesting relevance to processes such as invasion, metastasis-related migration, and tissue remodeling. These properties make DCBLD1 a useful target for mechanistic studies connecting surface receptor signaling to downstream pathway activation and cellular state transitions.

DCBLD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DCBLD1 expression without altering the underlying DNA sequence.

DCBLD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DCBLD1 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 DCBLD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DCBLD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DCBLD1 locus and enabling the study of DCBLD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DCBLD1 pathway restoration in tumor cells with silenced or reduced DCBLD1 expression.

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