BLOS1 CRISPR Activation Plasmid (h): sc-406825-ACT

BLOC1S1 encodes BLOS1, a core subunit of the biogenesis of lysosome-related organelles complex-1 (BLOC-1) that supports endosomal sorting and cargo trafficking required for normal lysosome-related organelle function. Through its role in vesicle maturation and membrane protein delivery, BLOS1 contributes to cellular processes including protein turnover, organelle homeostasis, and regulated secretion. Perturbation of BLOC-1 components has been linked to defects in intracellular trafficking pathways with relevance to neurodevelopmental and neuropsychiatric phenotypes, and to altered immune and pigment organelle biology. As a result, BLOC1S1 is frequently studied in models of endolysosomal dysfunction and pathway-level regulation of intracellular transport.

BLOS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BLOC1S1 expression without altering the underlying DNA sequence.

BLOS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BLOC1S1 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 BLOC1S1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BLOS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BLOC1S1 locus and enabling the study of BLOS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BLOS1 pathway restoration in tumor cells with silenced or reduced BLOC1S1 expression.

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