Exploiting the RUSH System to Study Lytic Granule Biogenesis in Cytotoxic T Lymphocytes

Abstract

The Retention Using Selective Hooks (RUSH) system allows for the synchronized release of one or more proteins of interest from a donor endomembrane compartment, usually the endoplasmic reticulum, and the subsequent monitoring of their traffic toward acceptor compartments. Here we describe the RUSH system applied to cytotoxic T cells to characterize the biogenesis of lytic granules, using as a proof-of-concept granzyme B trafficking to this specialized compartment.

Keywords: Confocal microscopy; Cytotoxic granules; Granzyme B; Intracellular trafficking; Lytic granule biogenesis; RUSH system; Spinning disk microscopy.