MAL CRISPR Activation Plasmid (h): sc-403024-ACT

Human MAL encodes a highly hydrophobic tetraspan proteolipid enriched in glycosphingolipid- and cholesterol-rich membrane microdomains, where it contributes to apical membrane trafficking, vesicular sorting, and stabilization of specialized membrane domains. MAL participates in polarized epithelial transport and is implicated in organization of lipid rafts that influence receptor localization and signal propagation at the plasma membrane. Its expression is lineage- and differentiation-dependent, making it a useful marker for epithelial and lymphoid cell states and for studying membrane compartmentalization. Dysregulated MAL expression has been reported across multiple cancer contexts and in epithelial remodeling, supporting research into how altered trafficking and raft dynamics relate to oncogenic signaling and cell polarity.

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

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

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