Cellular reprogramming of mammalian glia to an induced neuronal fate holds potential for restoring diseased brain circuits. While the proneural factor Ascl1 is widely used for neuronal reprogramming, in the early postnatal mouse cortex Ascl1 fails to induce glia-to-neuron conversion, instead promoting proliferation of oligodendrocyte progenitor cells (OPC). Since Ascl1 activity is post-translationally regulated, here we investigated the consequences of mutating six serine phospho-acceptor sites to alanine (Ascl1SA6) on lineage reprogramming in vivo. Ascl1SA6 exhibited increased neurogenic activity in glia of the early postnatal mouse cortex, an effect enhanced by co-expression of Bcl2. Genetic fate-mapping revealed that most induced neurons originated from astrocytes while only a few derived from OPCs. Intriguingly, many Ascl1SA6/Bcl2-induced neurons expressed parvalbumin and were capable of high-frequency action potential firing. Our study demonstrates authentic conversion of astroglia into neurons featuring subclass hallmarks of cortical interneurons, advancing our scope of engineering neuronal fates in the brain.

Reprogramming early cortical astroglia into neurons with hallmarks of fast-spiking parvalbumin-positive interneurons by phospho-site deficient Ascl1