Leveraging transposon traps to isolate cell cycle-specific reporter cells

Abstract We present an efficient method for isolating multiple reporter cell clones that are specifically expressed at different stages of the cell cycle, potentially encompassing every stage with staggered peaks in reporter expression. Following random transposon insertion of the GAL4FF-UAS-mediated Fluc-P2A-EGFP reporter system (Ishikawa et al., 2021.), we isolated numerous clones from the EGFP(+) fraction, which remained resistant to elimination even after repeated sorting of EGFP(−) cells. Upon treatment with nocodazole, thymidine, or imatinib to arrest cells at specific cell cycle phases, we observed varied increases or decreases in reporter expression among the clones. These results, along with the analysis of trapped gene sequences, indicate that the transposon system efficiently traps diverse cell cycle-related genes. Expanding this method and profiling gene expression timing at specific cell cycle phases holds promise for identifying new cell cycle phase-specific marker genes, or for screening and evaluating compounds with anti-cancer activity, thereby potentially contributing to drug development.