LRRK2 CRISPR Activation Plasmid (m): sc-426167-ACT

Leucine-rich repeat kinase 2 (LRRK2) is a large multidomain serine/threonine kinase and GTPase that coordinates vesicular trafficking, endolysosomal homeostasis, cytoskeletal dynamics, and autophagy through interactions with Rab GTPases and associated signaling networks. In mouse cells, Lrrk2 activity influences phosphorylation-dependent remodeling of membrane trafficking pathways, linking innate immune responses and mitochondrial quality control to cellular stress adaptation. Dysregulated LRRK2 signaling has been widely studied in neurodegeneration-associated mechanisms, including altered proteostasis, impaired lysosomal function, and aberrant inflammatory signaling. These features make Lrrk2 a useful node for dissecting pathway crosstalk between neuronal maintenance, glial activation, and intracellular transport.

LRRK2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Lrrk2 expression without altering the underlying DNA sequence.

LRRK2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Lrrk2 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 Lrrk2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRRK2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Lrrk2 locus and enabling the study of LRRK2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRRK2 pathway restoration in tumor cells with silenced or reduced Lrrk2 expression.

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