LC3B CRISPR Activation Plasmid (h): sc-417828-ACT

MAP1LC3B encodes microtubule-associated protein 1 light chain 3 beta (LC3B), a ubiquitin-like modifier that is cleaved and lipidated to form LC3-II on nascent autophagosomal membranes. LC3B functions as a core scaffold for autophagy by binding LIR-motif cargo receptors such as SQSTM1/p62, thereby coordinating selective cargo sequestration and autophagosome maturation within the lysosomal degradation pathway. This protein integrates nutrient and stress signaling, including MTOR and AMPK-regulated autophagy initiation, and is routinely used to monitor autophagic flux and vesicle dynamics. Dysregulated LC3B-associated autophagy has been implicated in contexts including neurodegeneration, infection, and cancer cell stress adaptation, supporting mechanistic studies of proteostasis and organelle quality control.

LC3B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAP1LC3B expression without altering the underlying DNA sequence.

LC3B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAP1LC3B 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 MAP1LC3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LC3B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAP1LC3B locus and enabling the study of LC3B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LC3B pathway restoration in tumor cells with silenced or reduced MAP1LC3B expression.

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