2021/07/06 by Katrin Mangold, J. Mašek, Jingyan He +3 · 1 voice · 1 citation
Biochemistry, Genetics and Molecular Biology · Neuroscience · #CRISPR and Genetic Engineering #Genetic Neurodegenerative Diseases #Pluripotent Stem Cells Research
paper · doi:10.1016/j.crmeth.2021.100043
openalex publication_date 2021/07/06 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/22
Genetic loss and gain of function in mice have typically been studied by using knockout or knockin mice that take months to years to generate. To address this problem for the nervous system, we developed NEPTUNE (NEural Plate Targeting by in Utero NanoinjEction) to rapidly and flexibly transduce the neural plate with virus prior to neurulation, and thus manipulate the future nervous system. Stable integration in >95% of cells in the brain enabled long-term overexpression, and conditional expression was achieved by using cell-type-specific MiniPromoters. Knockdown of Olig2 by using NEPTUNE recapitulated the phenotype of Olig2−/− embryos. We used NEPTUNE to investigate Sptbn2, mutations in which cause spinocerebellar ataxia type 5. Sptbn2 knockdown induced dose-dependent defects in the neural tube, embryonic turning, and abdominal wall closure, previously unreported functions for Sptbn2. NEPTUNE thus offers a rapid and cost-effective technique to test gene function in the nervous system and can reveal phenotypes incompatible with life.