Output Catalog
ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We’ve created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
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Output Type
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Program
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CRN Team Name
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Theme
Cryo-electron tomography of thinned synapses
Protocol for cryo-ET of thinned synapses in rat neuron cultures is detailed. Cryo-FIB milling creates thin samples for high-resolution imaging. Synapse targeting with and without cryo-FLM correlation is explained.
Preparing primary sandwich hippocampal neuron cultures for cryo-electron tomography
Sandwich hippocampal neuron cultures have low cell density and few glial cells, making them ideal for targeting neurons for cryo-electron tomography.
Single particle analysis of α-Synuclein fibrils
Aggregates of α-Synuclein are linked to Parkinson’s, multiple system atrophy, and Lewy body dementia. This protocol outlines near-atomic resolution structures of these amyloid fibrils using cryo-EM single particle analysis.
CRISPR tagging of the TMEM192 gene with 3xHA in H9 ES AAVS-NGN2 cells for Lyso-IP
Protocol for adding a 3X HA tag to TMEM192 gene in human cells using CAS9 and recombination template is described for H9 ES AAVS1-NGN2 cells.
Analysis of ER Flux in Cultured Induced Neurons using Keima ER reporters
Associated with preprint: https://doi.org/10.1101/2023.06.26.546565
Analysis of ER structures in Cultured Induced Neuron axons
Associated with preprint: https://doi.org/10.1101/2023.06.26.546565
Characterizing spatial and temporal properties of ER-phagy receptors
Associated with preprint: https://doi.org/10.1101/2023.06.26.546565
Microscopy-based measurements of p62 recruitment in HeLa
Associated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Microscopy-based pUb-coverage measurements of mitochondria in iNeurons
Associated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Evaluation of pUb kinetics using 3D-SIM
Associated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Lipidomic analysis of tissue culture cells, tissues, and purified organelles
Lipids are a class of molecules that have roles in energy storage, plasma membrane integrity, and signaling events. To gain more understanding of the functions and roles that lipids play in biology, researchers employ discovery analytical…
Immunohistochemistry of rTg4510 mouse brain
The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.
Lentivirus production for primary neuron transduction
This protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman…
Flow cytometry-based measurement of trafficking of Golgi to the lysosome via autophagy
A protocol was developed to measure Golgi trafficking to lysosomes through autophagy in HEK293 cells using Keima-YIPF3/4 reporters with flow cytometry.
Proximity biotinylation of ATG8 proteins and selective autophagy receptors
Autophagy forms autophagosomes around cellular cargo for degradation in lysosomes. Proximity biotinylation with APEX2 fused to ATG8 proteins identifies proteins near autophagic cargo receptors during selective autophagy activation by nutrient stress
