SorLA CRISPR Activation Plasmid (h): sc-403088-ACT

Human SORL1 encodes SorLA, a VPS10-domain sorting receptor that regulates endosomal trafficking and receptor recycling in neurons and other cell types. SorLA modulates intracellular routing of amyloid precursor protein (APP) and influences amyloidogenic processing by directing cargo between the trans-Golgi network, endosomes, and the cell surface. Through its role in vesicular transport and lysosome-associated turnover, SorLA contributes to proteostasis, synaptic function, and lipid receptor handling, linking SORL1 dysregulation to Alzheimer’s disease genetics and neurodegeneration-associated endosomal defects. Altered SorLA activity is therefore widely studied in pathways governing APP processing, endosome maturation, and neuronal homeostasis.

SorLA CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SORL1 expression without altering the underlying DNA sequence.

SorLA CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SORL1 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 SORL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SorLA expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SORL1 locus and enabling the study of SorLA-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SorLA pathway restoration in tumor cells with silenced or reduced SORL1 expression.

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