Molecular Genetic Identification of Skeletal Remains from the Unmarked Graves of the First Half of the 16th Century from the Necropolis of the Ascension Cathedral of the Moscow Kremlin

The necropolis of Russian grand princesses and tsarinas, buried in the period from 1407 to 1731 in the Ascension Cathedral of the Moscow Kremlin, suffered considerable damage, and many sarcophagi appeared to be unmarked, which led to the difficulties in identification of skeletal remains in them. The present study is focused on the identification of unidentified remains, presumably belonging to the descendants of the Grand Princess of Moscow Sophia Palaiologina (1455-1503), her daughter Evdokia Ivanovna, and granddaughter Anastasia Petrovna. Historical chronicles have preserved information on the burial of two female representatives of the princely family in this temple in the first half of the 16th century. Genetic analysis made it possible to successfully perform indirect identification of skeletal remains from two graves of the period of the Russian Middle Ages from the necropolis of the Ascension Monastery in the Moscow Kremlin and to establish relative relationships among the buried. Analysis of the polymorphic autosomal microsatellite loci, as well as hypervariable regions of the mitochondrial DNA (mtDNA) from three female skeletons, revealed their close relationships. Analysis of the mtDNA hypervariable loci showed that all three skeletons, including the remains of Grand Princess Sophia Palaiologina, belonged to the same mitogroup (HV0) which is the most common among European populations. The data obtained are extremely important for the historical reconstruction of the formation in the 16th century of one of the two most prestigious tombs of the Moscow Kremlin. In addition, the data of the present study can help in the reconstruction of family burial sites of the highest nobility of the medieval Russian state.