Gene editing for Spinocerebellar ataxia type 3 taking advantage of the human ATXN3L paralog as replacement gene

Human embryonic kidney 293 T (HEK293T) cells

HEK293T cells (mycoplasma-negative, ATCC, LGC Standards GmbH, Wessel, Germany) were cultured in DMEM-Glutamax supplemented with 10% FBS and 1% penicillin/streptomycin (Gibco, Life Technologies, Zug, Switzerland) at 37 °C under an atmosphere containing 5% CO₂. Cells were passaged twice weekly after trypsin treatment for dissociation (Gibco, Life Technologies, Zug, Switzerland) and plated at a density of 2 × 10⁶ cells/cm² in T175 flasks.

Neuronal progenitor cells culture and transduction

The…