CRISPR-mediated Loss of Function Analysis in Cerebellar Granule Cells Using <em>In Utero</em> Electroporation-based Gene Transfer

Somatic gene delivery for flexiblein vivomodeling of high-risk sarcoma

“…We substantially optimized muscle electroporation in P30 mice compared to previous studies (Huang et al, 2017; Young and Dean, 2015), but also adapted the procedure to apply it for the first time in neonatal mice, marking the earliest murine soft-tissue electroporation to date. Of note, in utero EPO can be applied to brain tissue, but relies on injections into the lumen of cerebral ventricles (Feng et al, 2018), a structural feature not present in other tissues. In assessing the conditions for the highest transfection efficiency of muscle tissue at two postnatal developmental time points, we increased the probability of hitting cells permissive for transformation even if they are rare.…”

“…Similarly, TFCP2-fused RMS typically occur intraosseusly or are bone-associated, suggesting a non-myogenic origin (Schöpf et al, 2024). For some entity-specific alterations, embryonic cell states may be required for oncogenic transformation, as previously described for induction of brain tumors by in utero electroporation (IUE) (Feng et al, 2018). This likely explains why Smarcb1 -inactivation alone was not sufficient for tumorigenesis in our model system, and concomitant Trp53 - inactivation was required to drive EpS/MRT-like tumors (Han et al, 2016; Li et al, 2021).…”