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

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 aggre-gation in Huntington's disease cellular models. The protocol can be readily repur-posed 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

Automated summary

In this article, a protocol for preparing small extracellular vesicles (sEVs) enriched in the chaperone protein DNAJB6 using genetic engineering techniques is presented. The steps to prepare cell lines overexpressing DNAJB6, as well as the isolation and characterization of sEVs from cell conditioned media, are described. Additionally, assays to examine the effects of DNAJB6-loaded sEVs on protein aggregation in Huntington's disease cellular models are discussed. The protocol can be applied to study protein aggregation in other neurodegenerative disorders or extended to investigate other therapeutic proteins. For complete details on the use and execution of this protocol, please refer to Joshi et al. (2021).