mucolipin 1 CRISPR Activation Plasmid (h): sc-403931-ACT

MCOLN1 encodes mucolipin 1 (TRPML1), an endolysosomal cation channel that regulates Ca²⁺ release required for membrane trafficking, lysosome biogenesis, and autophagosome–lysosome fusion. TRPML1 integrates with lysosomal signaling networks involving PI(3,5)P₂ and TFEB/TFE3-dependent transcriptional programs that coordinate cellular clearance and nutrient sensing. Disrupted MCOLN1 function perturbs endolysosomal homeostasis, leading to abnormal storage material accumulation and altered organelle dynamics characteristic of lysosomal storage disorders. As a result, MCOLN1 is frequently studied in pathways linking lysosomal Ca²⁺ signaling to autophagy, inflammation, and cellular stress responses in relevant cell models.

mucolipin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MCOLN1 expression without altering the underlying DNA sequence.

mucolipin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MCOLN1 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 MCOLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mucolipin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MCOLN1 locus and enabling the study of mucolipin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mucolipin 1 pathway restoration in tumor cells with silenced or reduced MCOLN1 expression.

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