The insight gained is impossible with classical biochemical techniques, as individual replisomes
are observed in real time rather than measuring an average of a population.
To this end, I will combine various cutting edge methods, like nanobody-based sequential purifications, SILAC mass spectrometry and 3D-SIM microscopy, to develop assays to isolate active replisomes
from human cells and study the identified factors mechanistically.
Specifically, we will investigate how RNA polymerases, replicative helicases, and replisomes
can be derailed or stalled.