SUMMARY OF RESEARCH DIRECTIONS –LABORATORY OF VIRAL AND CELLULAR GENETICS, INSTITUTE OF MOLECULAR GENETICS

0567914 - ÚMG 2023 RIV CZ eng A - Abstrakt
Hejnar, Jiří
SUMMARY OF RESEARCH DIRECTIONS –LABORATORY OF VIRAL AND CELLULAR GENETICS, INSTITUTE OF MOLECULAR GENETICS.
Czech Chemical Society Symposium Series. Roč. 20, č. 6 (2022), č. článku L-27. ISSN 2336-7202.
[Annual meeting of the National Institute of Virology and Bacteriology (NIVB) /1./. 30.11.2022-02.12.2022, Kutná Hora]
Grant CEP: GA MŠMT(CZ) LX22NPO5103
Institucionální podpora: RVO:68378050
Klíčová slova: avian leukosis virus * chicken * gene editing
Obor OECD: Virology
http://www.ccsss.cz/index.php/ccsss/issue/view/37/67

Avian leukosis virus (ALV) is a virus complex diversified through the virus-host coevolution (virus-host arms race) into several subgroups, each of which recognizes different cell surface receptor. In our laboratory, we employ this unique example of coevolution to study processes of virus adaptation, broadening the host range, and heterotransmission (in general, all zoonotic viruses arose in a process of heterotransmission). This knowledge could help in artificial creation of resistant animals using the CRISPR/Cas9 technology of gene editing in chicken, the natural host of ALV. Receptor alleles bearing simple substitutions of critical amino-acids at the virus binding domains (e.g., the recently prepared resistance to ALV-J subgroup1) are a good material for studying the escape mutations of the virus adapted to the new versions of receptors.Antiviral restriction factors and virus dependence factors are other subjects of virus-host coevolution and bidirectional evolutionary approach is necessary to characterize viral adaptations to new host species during the process of heterotransmission and zoonotic events as well as the changes in relevant host genes. Many host cell-encoded anti-HIV-1 activities already have been characterized in human cells. In our laboratory, we focus on the effects of chicken tetherin against avian viruses2. Chicken tetherin will be the primary target to genetic knock out or gene editing in vivo using the CRISPR/Cas9 with the final goal to demonstrate its role in aresilience to chicken diseases.The evolutionary approach enables the characterization of viral adaptations to new host species as well as adaptations necessary for long-term coevolution with hosts. These might include the appearance of novel antiviral activities of existing genes, the emergence of novel genes, or in some cases evolutionary losses of genes. In this respect, we focus on innate immunity factors in chicken, which exert antiviral effects, particularly against avian leukosis viruses, avian influenza, avian coronaviruses, etc. Chicken RIG-I and MDA-53are studied with the ultimate goal to edit their genes in vivo using the CRISPR/Cas9.All three aforementioned research directions will profit of our original technique of transgenesis and gene editing in chicken (Fig. 1), which consist of primordial germ cell (PGC) manipulation by CRISPR/Cas9 and orthotopic transplantation of PGCs into sterilized roosters4. In our laboratory, we also stdy cell-to-cell fusion during placenta development, which is critical for the proper formation of its outer layer called syncytiotrophoblast. Fusion of mononuclear cytotrophoblast and formation of the multinuclear syncytium is largely dependent on the human proteins called Syncytins. Syncytin-1 and Syncytin-2, the retroviral envelope glycoproteins of two distinct human endogenous retroviruses. Using our original heterologous ectopic system of retrovirus-receptor interaction and dual luciferase cell fusion technique5, we will analyze contribution of syncytin-1/2 and receptors hASCT1/2 for trophoblast fusion and identify amino-acids critical for syncytin-receptor binding. CryoEM structural analysis will be performed to support the functional analyses. In parallel, epigenetic repression of syncytins in non-placental tissues will be studied. This projet might result in identification of mutations behind the human idiopathic infertility.
Trvalý link: https://hdl.handle.net/11104/0339179