Project description:throughput profiling of WT1 in M15 mouse mesonephric cells and M15 genome edited for the KTS isoforms. The genome-edited lines were derived from the M15 cells by transfection and puromycin selection of cells with the guide RNA expressing plasmids. Briefly, the guide RNAs were designed against the splice site that is essential for KTS isoform. Out of the designed guide RNAs, some of them were cloned into the pX601 vector with the Sa Cas9 variant, and clone confirmation was done using sequencing. Isoform-specific lines were established by providing oligos for repair which represented the region corresponding to the codons coding for KTS or not.
