1,721,142 research outputs found
Genome editing technologies to treat rare liver diseases
Full text linkLiver has a central role in protein and lipid metabolism, and diseases involving hepatocytes have often repercussions on multiple organs and systems. Hepatic disorders are frequently characterized by production of defective or non-functional proteins, and traditional gene therapy approaches have been attempted for years to restore adequate protein levels through delivery of transgenes. Recently, many different genome editing platforms have been developed aimed at correcting at DNA level the defects underlying the diseases. In this Review we discuss the latest applications of these tools applied to develop therapeutic strategies for rare liver disorders, in particular updating the literature with the most recent strategies relying on base editors technology
SOME THEORETICAL ASPECTS OF THE MECHANISM OF ACTION OF BENZHYDRAZONE, AN INHIBITOR OF GLYCOSYLATION IN HERPES-SIMPLEX VIRUS
No full textSome hypotheses of the mechanism of action of benzhydrazone, a selective inhibitor of herpes simplex virus glycoprotein biosynthesis, are presented in this paper. Comparative analysis of different models indicates that the drug may interfere with herpes simplex virus DNA. Thus, benzhydrazone seems to act like a bifunctional alkylating agent
Live-cell analysis of human cytomegalovirus dna polymerase holoenzyme assembly by resonance energy transfer methods
Full text linkHuman cytomegalovirus (HCMV) genome replication is a complex and still not completely understood process mediated by the highly coordinated interaction of host and viral products. Among the latter, six different proteins form the viral replication complex: a single-stranded DNA binding protein, a trimeric primase/helicase complex and a two subunit DNA polymerase holoenzyme, which in turn contains a catalytic subunit, pUL54, and a dimeric processivity factor ppUL44. Being absolutely required for viral replication and representing potential therapeutic targets, both the ppUL44–pUL54 interaction and ppUL44 homodimerization have been largely characterized from structural, functional and biochemical points of view. We applied fluorescence and bioluminescence resonance energy transfer (FRET and BRET) assays to investigate such processes in living cells. Both interactions occur with similar affinities and can take place both in the nucleus and in the cytoplasm. Importantly, single amino acid substitutions in different ppUL44 domains selectively affect its dimerization or ability to interact with pUL54. Intriguingly, substitutions preventing DNA binding of ppUL44 influence the BRETmax of protein–protein interactions, implying that binding to dsDNA induces conformational changes both in the ppUL44 homodimer and in the DNA polymerase holoenzyme. We also compared transiently and stably ppUL44-expressing cells in BRET inhibition assays. Transient expression of the BRET donor allowed inhibition of both ppUL44 dimerization and formation of the DNA polymerase holoenzyme, upon overexpression of FLAG-tagged ppUL44 as a competitor. Our approach could be useful both to monitor the dynamics of assembly of the HCMV DNA polymerase holoenzyme and for antiviral drug discovery
Virus-based vectors for gene expression in mammalian cells: Retrovirus
No full textHigh-level expression of proteins in animal cells has been very informative in studies of protein and cellular function and in many cases has relied on virus-derived expression vectors. Many viruses have evolved to maximize expression of their proteins in host cells and are therefore a good starting point for construction of efficient expression vectors. Retroviruses are diploid, single-stranded, positive-sense RNA viruses [1]. As an obligate step of the retrovirus life cycle, the RNA genome is
converted into DNA and then integrated into the host cell chromosome in the form of provirus. The provirus replicates as the host cell chromosome replicates and is transmitted to all progeny cells. This ability of retroviruses to stably introduce new
genetic information into the target cells led to the development of retroviruses as vehicles for the stable transfer of genes [2–4]. Indeed, retroviral vectors have been used for a variety of experimental applications, including insertional mutagenesis, cell
lineage studies, the creation of transgenic animals and the expression of foreign genes into mammalian cells, both in vitro and in vivo as documented by the majority of gene therapy clinical trials. An ideal vector should guarantee not only high efficient gene transfer but also appropriately regulated and stable gene expression from a safely integrated provirus. Currently, efforts are devoted to achieve these goals. This
chapter focuses on recent progress on retroviral vector design and applications
Exposure to bacterial cell wall products triggers an inflammatory phenotype in hepatic stellate cells.
No full text
Effect of coumermycin on influenza A virus replicative cycle.
No full textAbstract
Coumermycin A1, a Novobiocin related antibiotic, has been tested in vivo and in vitro for its activity on Influenza A viruses. In a range of concentrations between 3 and 5 micrograms/ml virus production was drastically reduced. The drug was able to inhibit virus release into culture medium also if added up to the sixth hour following infection and its action was reversible within this time. The synthesis of virus induced polypeptides was generally depressed but production of the HA was more deeply inhibited. Viral transcriptase activity in vitro was also affected by the presence of Coumermycin A1 but at doses which seem to high to consider this event as a phenomenon likely to play a role in vivo. It is suggested that the antiviral activity of the drug is mediated by the inhibition of the host cell metabolism.
PMID: 6547762 [PubMed - indexed for MEDLINE
- …
