Preparation of chaperone-loaded neural stem cell-derived extracellular vesicles to reduce protein aggregation in Huntington's disease cellular models

STAR Protoc. 2023 Mar 17;4(1):102134. doi: 10.1016/j.xpro.2023.102134. Epub 2023 Feb 25.

Abstract

Here, we present a protocol using genetic engineering techniques to prepare small extracellular vesicles (sEVs) enriched in the chaperone protein DNAJB6. We describe steps to prepare cell lines overexpressing DNAJB6, followed by the isolation and characterization of sEVs from cell conditioned media. Further, we describe assays to examine effects of DNAJB6-loaded sEVs on protein aggregation in Huntington's disease cellular models. The protocol can be readily repurposed to study protein aggregation in other neurodegenerative disorders or extended to other therapeutic proteins. For complete details on the use and execution of this protocol, please refer to Joshi et al. (2021).1.

Keywords: Cell Biology; Cell Separation/Fractionation; Gene Expression; Health Sciences; Microscopy; Molecular Biology; Molecular/Chemical Probes; Neuroscience; Protein Biochemistry; Stem Cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Extracellular Vesicles* / metabolism
  • HSP40 Heat-Shock Proteins / genetics
  • HSP40 Heat-Shock Proteins / metabolism
  • Humans
  • Huntington Disease* / genetics
  • Huntington Disease* / metabolism
  • Huntington Disease* / therapy
  • Molecular Chaperones / genetics
  • Molecular Chaperones / metabolism
  • Nerve Tissue Proteins / metabolism
  • Neural Stem Cells* / metabolism
  • Neurodegenerative Diseases*
  • Protein Aggregates

Substances

  • Protein Aggregates
  • DNAJB6 protein, human
  • Nerve Tissue Proteins
  • Molecular Chaperones
  • HSP40 Heat-Shock Proteins