|Differential NOVA2-Mediated Splicing in Excitatory and Inhibitory Neurons Regulates Cortical Development and Cerebellar Function.
|Year of Publication
|Saito, Y, Yuan, Y, Zucker-Scharff, I, Fak, JJ, Jereb, S, Tajima, Y, Licatalosi, DD, Darnell, RB
|2019 Feb 20
RNA-binding proteins (RBPs) regulate genetic diversity, but the degree to which they do so in individual cell types in vivo is unknown. We developed NOVA2 cTag-crosslinking and immunoprecipitation (CLIP) to generate functional RBP-RNA maps from different neuronal populations in the mouse brain. Combining cell type datasets from Nova2-cTag and Nova2 conditional knockout mice revealed differential NOVA2 regulatory actions on alternative splicing (AS) on the same transcripts expressed in different neurons. This includes functional differences in transcripts expressed in cortical and cerebellar excitatory versus inhibitory neurons, where we find NOVA2 is required for, respectively, development of laminar structure, motor coordination, and synapse formation. We also find that NOVA2-regulated AS is coupled to NOVA2 regulation of intron retention in hundreds of transcripts, which can sequester the trans-acting splicing factor PTBP2. In summary, cTag-CLIP complements single-cell RNA sequencing (RNA-seq) studies by providing a means for understanding RNA regulation of functional cell diversity.
|R01 NS081706 / NS / NINDS NIH HHS / United States
UM1 HG008901 / HG / NHGRI NIH HHS / United States
R01 NS034389 / NS / NINDS NIH HHS / United States
R35 NS097404 / NS / NINDS NIH HHS / United States
R01 GM107331 / GM / NIGMS NIH HHS / United States
R56 NS034389 / NS / NINDS NIH HHS / United States