Combined U-Pb, O and Hf isotope data in zircon allows discrimination between juvenile and reworked crust, and is therefore a useful tool for understanding formation and evolution of the continental crust. The crustal evolution of basement rocks in central North America (Laurentia) is poorly constrained, as it is almost entirely overlain by Palaeozoic cover. In order to improve our understanding of the evolution of this region we present U-Pb, O and Hf isotope data from zircon in drill-core samples from the subsurface basement of Ohio. The Hf isotope data suggests juvenile crust formation at ~1650 Ma followed by continued reworking of a single reservoir. This 1650 Ma reservoir was tapped at ~1450 Ma during the formation of the Granite-Rhyolite Province and subsequently reworked again during the Grenvillian orogeny. The ~1650 Ma crust formation model age for the suite of samples along with the presence of ~1650 Ma magmatic rocks suggests an eastward extension of the Mazatzal Province (or Mazatzal-like crust) and makes it a possible protolith to the subsurface basement of Ohio and surrounding Mesoproterozoic (i.e. Grenville-age) rocks. The eastward extension of this ~1650 Ma crustal reservoir into Ohio requires a revision of the crustal boundary defined by Nd isotopic data to be located further east, now overlapping with the Grenville front magnetic lineament in Ohio. In fact, the easternmost sample in this study is derived from a more depleted reservoir. This limits the extent of >1.5 Ga basement in subsurface Ohio and constrains the location of the crustal boundary. Further, syn-orogenic magmatism at ~1050 Ma suggests a potential extrapolation of the Interior Magmatic Belt into Ohio. Oxygen isotopic data in zircon suggests that during Grenvillian metamorphism, zircon recrystallisation occurred in the presence of heavy δ18O fluids resulting in zircon with elevated δ18O values.