A genome editing approach to the study of Parvovirus B19

Parvovirus B19 (B19V) is a ssDNA virus, with a 5596 nt long genome encapsidated within an icosahedral capsid with a diameter of 22 nm. Viral proteins are subdivided into structural and non-structural: the main non-structural one is the NS1, while the 2 structural proteins VP1 and VP2 assemble originating the capsid shell. B19V tropism is mainly limited to erythroid progenitor cells (EPCs), however, virus can be detected in several districts persisting in tissues possibly lifelong. The virus can induce anemia and erythroid aplasia. Therapeutic strategies are only symptomatic, so the search for antivirals is strongly active, with screenings showing the activity in vitro of different compounds like hydroxyurea, cidofovir and brincidofovir. In the first project, a functional minigenome of B19V was developed, able to express only the NS1 protein. This minigenome proved able to replicate and express the NS1 at levels comparable to unmodified clones. Furthermore, the ability of this minigenome to complement the function of NS1-deficient genomes was demonstrated, thus providing a proof-of-concept of B19V genome editing possibility and, at the same time, a useful tool to study the NS1 protein also as an antiviral target. In the second project I addressed the interplay between B19V and the cellular restriction factor APOBEC3B (A3B), a cytidine deaminase acting on ssDNA, whose footprint on B19V genome was proved by a bioinformatic sequence analysis performed by the hosting lab. To understand whether A3B still exerts activity and a potential antiviral effect on B19V, the UT7/EpoS1 cells were transduced with lentiviral vectors to silence A3B expression, then used as a model to study viral behavior. No significant role of A3B on B19V was demonstrated, in agreement with the hypothesis of viral adaptation to this cellular restriction factor; anyway, virus ability to alter A3B expression would deserve further investigations.

The book of Qohelet. A digital scholarly edition of the hebrew text

The objective of the present dissertation is a born-digital critical edition of the Hebrew Old Testament book of Qohelet. The edition is based on an extensive collation of variant readings from indirect sources – the Septuagint, the Peshitta, the works of St. Jerome (the Vulgate and the Commentary), and the Targum – as well as from direct sources such as the Qumran fragments and Hebrew medieval manuscripts. The ultimate goal of the edition is (a) to reproduce the earliest textual form, the Archetype, that can be reconstructed on the basis of the available evidence; and (b) to propose a rehabilitation of the Original of the Author by resorting, when necessary, to conjectural emendation. We date the Archetype to the II century BCE, corresponding to the date of Hebrew fragments from Qumran, while we place the Original between the V and III centuries BCE. Unlike previous critical editions of Qohelet, ours follows the so-called eclectic model, which involves the reconstitution of a critical text and the preparation of an apparatus of secondary variants. Our edition includes, moreover, new data, taken both from primary literature, such as the recently published Göttingen Septuagint, and from up-to-date studies and critical commentaries on the text of Qohelet. The work is made up of five main parts: an introduction, which sets forth the rationale of the edition and the methodology adopted; the collation, where the variants are listed in their original language; the commentary, where they are extensively discussed; the critical text accompanied by the apparatus, which presents a selection of authentic Hebrew variants taken from the collation; and finally, a translation of the critical text. The edition uses the mark-up language of the Text Encoding Initiative (TEI). It is realized in pdf, via LaTeX, and will be available in digital form, via the TEI-Publisher editor.