Using ATR and WEE1 inhibitors to induce origin firing
Inhibition of ATR or WEE1 kinase activity results in excessive dormant origin firing in S-phase cells through hyperactivation of CDK1 and CDK2 kinases. This gives us a unique opportunity to study replication initiation.
Using split-TurboID for proteomic screens
TurboID is an optimized biotin ligase for proximity labelling, and splitting it in two inactive halves of the protein allows us to study the formation of replication complex and the transient components involved in this process.
Using auxin-inducible degrons for rapid knockdowns to replication proteins
Replication proteins are notoriously difficult to study, largely due to the fact that their knockouts are lethal, and knockdowns have severe effects on the cell cycle. Transient knockdowns with mAID system allows us to study the roles of these proteins more easily.
We are using a combination of proteomic approaches to identify new players in human replication initiation.
We are studying the roles of non-catalytic replisome components in replication complex assembly and origin firing.
We are identifying the specific roles and substrates of cell cycle kinases in S-phase entry and replication initiation.