PLX322254

GSE98838: Combining RNA-Seq and Somatic CRISPR Mutagenesis to Study Mouse Neural Development in vivo

• Organsim mouse
• Type RNASEQ
• Target gene
• Project ARCHS4

We used RNA sequencing to identify candidate regulators of interactions between photoreceptor axons and bipolar cell (BCs) dendrites in developing mouse retina. We chose three time points: P7, just after the OPL forms and synaptogenesis with BCs begins; P13, as synaptogenesis nears completion and sublamination begins; and P30, when the OPL is mature. We purified cone and rod photoreceptors separately by fluorescence activated cell sorting (FACS) using transgene markers: Rhoicre;Ai9 for rods and HRGPcre;Ai9 for cones. We purified ON BCs, which include ON cone bipolars plus rod bipolars using Grm6:GFP. As appropriate transgenic lines to separate RBCs from CBCs were not available, we performed RNAseq on cells from Grm6:GFP mice that were fixed and immunostained prior to FACS, allowing us to purify RBCs (GFP+PKC+) and CBCs (GFP+PKC-) from the same retinas. As PKC is not highly expressed at P7, profiling of developing rod bipolars separate from developing ON cone bipolars was restricted to P13. SOURCE: S Lawrence Zipursky University of California, Los Angeles