Studying Inner ear development through the use of the CARLIN mouse model to label progenitor cells, identifying the potency of early cell types in the otocyst

This experiment focuse on utilizing the CARLIN mouse model. The CARLIN system (CRISPR Array Repair Lineage tracing IN situ) is a genetic barcoding and lineage tracing tool developed to track the lineage and fate of individual cells in vivo over time. It was designed to allow researchers to reconstruct cell lineages and study progenitor cell behavior, especially in complex tissues like the hematopoietic system, the brain, and during development. Utilizing an inducible Cas9, it allows for temporal control of marking cells with unique heritable barcodes For this experiment, timed matings were performed, and plugs were checked the next morning. Doxycycline injection were performed on the pregnant dams at E7.5 and 8.5. The pups were collected for dissection at P4, 2-weeks post injections. Blunt dissection to remove and isolate the inner ear was performed, followed by single cell dissociation and EpCAM-focused FACS. Then, library prep of the collected cells were performed, and both bulk and single-cell sequencing was performed on this sample.