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  • Mechanisms Controlling Selective Elimination of Damaged Lysosomes

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    Lysophagy, triggered by membrane rupture, involves galectins binding to lysosomal contents to promote autophagic recycling. Damaged lysosomes are ubiquitylated, leading to autophagosome formation for degradation in healthy lysosomes.

  • pBMN_HA-TBK1 (kinase dead – D135N)

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    Plasmid: Used for the expression of TBK1 carrying a kinase dead mutation D135N (SMC Internal No. 1997).

  • pBMN_HA-TBK1 (E696K)

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    Plasmid: Used for the expression of TBK1 carrying E696K mutation (SMC Internal No. 1998).

  • Vesicular dysfunction and pathways to neurodegeneration

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    In this review, the pathways that have emerged as critical for neuronal survival in the human brain are discussed, illustrating the diversity of proteins and cellular events with three molecular case studies from different neurological diseases.

  • In Vivo Electrophysiology (Mouse)

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    This protocol describes the in vivo electrophysiology method for recording neuronal activity in mice.

  • Single cell survival assay

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    This protocol describes the experimental procedure used to measure single cell survival rates post nucleofection of human pluripotent stem cells (hPSCs). General Notes: 1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.

  • Immunohistochemistry

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    This protocol describes immunohistochemical staining of fixed brain sections.

  • RNAScope in situ hybridization (ISH) multiplex fluorescent

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    Instructions to perform ISH using RNAscope kit from ACDBio company optimized for frozen mouse brain and nodose ganglia sections.

  • pCAG-MBP-ATG9 (1-830)

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    Plasmid: Expresses human ATG9 C-terminal truncation mutant in mammalian cells.

  • Culture and transfection of HEK293T cells

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    This protocol describes a standard procedure culturing and transfecting HEK293T cells Protocol overview: A. Culturing HEK293T cells B. Transfection of HEK293T cells with Lipofectamine 2000

  • Staining of cells with GolgiTracker for Golgi flow cytometry analysis

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    Method that allows the staining of intact Golgi using GolgiTracker for subsequent flow cytometry analysis.

  • Highly efficient generation of isogenic pluripotent stem cell models using prime editing

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    Prime editing (PE) simplifies creating human pluripotent stem cell (hPSC) disease models by optimizing mRNA delivery with editing efficiency increased up to 13-fold, enabling correction or introduction of Parkinson's disease mutations in hPSCs.

  • Px330-EGFP-LRRK2-CRISPR/Cas9

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    It can be used to introduce LRRK2-G2019S mutation using CRISPR/Cas9 based genome editing. 

  • Single cell analysis of iPSC-derived midbrain organoids

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    The following script was used for analysis of gene corrected (GC) versus GBA1 mutant (MUT) midbrain organoids. The purpose was to combine, filter, integrate, and identify clusters and differentially expressed genes sets. This is part of a Collection of protocols (dx.doi.org/10.17504/protocols.io.8epv593dng1b/v1) for the paper "Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function" (https://doi.org/10.21203/rs.3.rs-1521848/v1)

  • pGST2-GST-TEV-OPTN S177D S473D

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    Plasmid for the expression and purification of GST-TEV-OPTN S177D S473D. Internal Ref: SMC1588

  • Midbrain organoid differentiation in spinner flasks

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    Midbrain differentiation protocol using spinner flasks

  • Rab8a expression and purification

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    Recombinant Rab8a expression and purification protocol as used by the Leschziner and Reck-Peterson Labs

  • Assaying starvation-induced autophagy in HeLa cells

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    A method for assaying starvation-induced autophagy in HeLa cells that have been transfected with Halo-tagged constructs.

  • pCAG-ATG13 (363-517)-ULK1 (836-1050)-TSF

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    Plasmid for mammalian expression of ATG13 C-terminal and ULK1 MIT domain fusion complex.

  • Subcellular and regional localization of mRNA translation in midbrain dopamine neurons

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    Using, highly sensitive ribosome-bound RNA sequencing and imaging to characterize the translatome, the authors uncovered local mRNA translation of dopamine synthesis, release, and reuptake machinery in dendrites, but not axons.

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