Engineered Aim-Based Selective Autophagy to Degrade Proteins and Organelles

Abstract

Techniques for disrupting of protein function are essential for biological researches and therapeutics development. Though well-established, genetic perturbation strategies may have off-target effects and/or trigger compensatory mechanisms, and cannot efficiently eliminate existing protein variants or aggregates. Therefore, precise and direct protein-targeting methods are highly desired. Here we describe a novel method for targeted protein clearance by engineering an autophagy receptor with a binder to provide target specificity and an ATG8-binding motif (AIM) to link the targets to nascent autophagosomes, thus harnessing the autophagy machinery for degradation. We demonstrate its specificity and broad potentials by degrading various fluorescent-tagged proteins and peroxisome organelle, using a tobacco-based transient expression system, and by degrading endogenous proteins in transgenic Arabidopsis expressing engineered receptors. With the wide substrate scope and specificity of selective autophagy, our method provides a convenient and robust strategy for eliminating proteins and aggregates, and may enable developing new treatments for protein-related disorders.

Emerging degrader technologies engaging lysosomal pathways

“…Based on AIM/LIR-LC3 binding knowledge, Luo et al designed an AIM/LIR based selective degrader that can precisely degrade target proteins. 74 They designed a degrader composed of a target-specific binder and an AIM/LIR to tether the protein of interest (POI) to nascent autophagosomes. The system was shown to degrade various fluorescence-tagged proteins and peroxisome organelles.…”