Profiling the transcriptome of layer 5 intratelencephalic-projection neurons (IT-PNs) in the mature mouse motor cortex

Low input RNA-seq analysis of fluorescently activated cell sorted IT-PNs from layer 5 of the mature mouse motor cortex. Comparison of two distinct IT-PN subtypes previously described by Tantirigama et al. (2014) identifies unique gene expression. Overall design: A Fezf2-Gfp reporter mouse model was injected at postnatal day 23 - 25 with the retrograde tracer, cholera toxin subunit b conjugated to Alexa 647, in M1 of the left hemisphere (n = 5). Five to seven days after injection, the layer 5 of the right hemisphere M1 was dissected and dissociated for FACS-purification of Fezf2+ and Fezf2- IT-PNs. A novel approach to low-input RNA-seq was used to generate cDNA libraries before sequencing.