Abstract: In the almost seven decades since the initial discovery of the lysosome as an organelle, our understanding of the role of lysosomes has greatly evolved. We now know lysosomal function encompasses far more than its traditionally described role as the cell’s “garbage disposal”, referring to its well-established catabolic function. Lysosomes are integral to maintaining cellular health and viability, and they act as a major signaling hub within the cell (Ballabio and Bonifacino, 2020). Lysosomes regulate autophagy, a key mechanism for regulating cellular homeostasis. The aberrant regulation of different lysosomal pathways is frequently observed in neurodegenerative diseases. Studies have identified a multitude of lysosomal genes that are now implicated in disorders including Parkinson’s disease (PD), Alzheimer’s disease (AD), and amyotrophic lateral sclerosis (Udayar et al., 2022). For example, many genes associated with late-onset AD, such as APOE and PLD3, are associated with autophagy-lysosomal pathways (Van Acker et al., 2019). In addition, lysosomal dysfunction contributes to the aggregation of alpha-synuclein in PD and the build-up of beta-amyloid and tau-containing neurofibrillary tangles in AD (Mazzulli et al., 2016; Feng et al., 2020). However, the development of therapeutics targeting PD, AD, and related neurodegenerative diseases has been hindered by the fact that we still do not fully understand the cellular and molecular changes contributing to lysosomal dysfunction in neurodegeneration. Thus, an emphasis has been placed on developing new techniques that can elucidate minute alterations in cellular and molecular processes. Over the past few years, both the development of methods for the rapid isolation of intact lysosomes and new CRISPR/Cas9-based genetic screens have furthered our knowledge of many lysosomal functions. Recently, studies with isolated lysosomes have shown the dynamic nature of nutrient exchange between the lysosome and the cytosol (Abu-Remaileh et al., 2017). In parallel, the first CRISPR interference (CRISPRi)-based genetic screens in human induced pluripotent stem cell (iPSC)-derived neurons have yielded novel insights into the relationship between glycosphingolipid accumulation and elevated oxidative stress (Abu-Remaileh et al., 2017; Tian et al., 2021). These findings pave the way for expanding our knowledge on how lysosomal dysfunction relates to neurodegeneration (Figure 1).