Novel astrocytic translatome isolation pipeline uncovers regional and sex-specific differences in mouse brain cortex
Vsevolod Treshin, Despoina Binou, Madlen Haase, Ina Ingrisch, Anja Urbach, Jean-Christopher Hennings, Martin Bens, Otto W Witte, Sidra Gull, Silvio Schmidt · FENS Forum 2024
Astrocytes, with their intricate morphology, pose challenges for cell-specific RNA isolation using traditional methods, which often result in loss of RNA localised in distal processes. We tackle this issue by adopting the RiboTag method (Sanz et al 2009) in RPL22tm1.1Psam mice. This technique isolates ribosome-associated mRNA (translatome) cell-specifically from crude tissue lysates through ribosomal HA tagging via cell-type specific Cre expression. Despite the widespread use of RiboTag, its detailed adaptation for complex cells like astrocytes, especially in high-throughput scenarios, remains
ePoster · Neuroscience