Due to the fast global spreading of the Severe Acute Respiratory Syndrome Coronavirus - 2 (SARS-CoV-2), prevention and treatment options are direly needed in order to control infection-related morbidity, mortality, and economic losses. Although drug and inactivated and attenuated virus vaccine development can require significant amounts of time and resources, DNA and RNA vaccines offer a quick, simple, and cheap treatment alternative, even when produced on a large scale. The spike protein, which has been shown as the most antigenic SARS-CoV-2 protein, has been widely selected as the target of choice for DNA/RNA vaccines. Vaccination campaigns have reported high vaccination rates and protection, but numerous unintended effects, ranging from muscle pain to death, have led to concerns about the safety of RNA/DNA vaccines. In parallel to these studies, several open reading frames (ORFs) have been found to be overlapping SARS-CoV-2 accessory genes, two of which, ORF2b and ORF-Sh, overlap the spike protein sequence. Thus, the presence of these, and potentially other ORFs on SARS-CoV-2 DNA/RNA vaccines, could lead to the translation of undesired proteins during vaccination. Herein, we discuss the translation of overlapping genes in connection with DNA/RNA vaccines. Two mRNA vaccine spike protein sequences, which have been made publicly-available, were compared to the wild-type sequence in order to uncover possible differences in putative overlapping ORFs. Notably, the Moderna mRNA-1273 vaccine sequence is predicted to contain no frameshifted ORFs on the positive sense strand, which highlights the utility of codon optimization in DNA/RNA vaccine design to remove undesired overlapping ORFs. Since little information is available on ORF2b or ORF-Sh, we use structural bioinformatics techniques to investigate the structure-function relationship of these proteins. The presence of putative ORFs on DNA/RNA vaccine candidates implies that overlapping genes may contribute to the translation of smaller peptides, potentially leading to unintended clinical outcomes, and that the protein-coding potential of DNA/RNA vaccines should be rigorously examined prior to administration.
- MeSH
- DNA vakcíny škodlivé účinky genetika MeSH
- glykoprotein S, koronavirus genetika MeSH
- kodon MeSH
- konformace nukleové kyseliny MeSH
- lidé MeSH
- messenger RNA MeSH
- mRNA vakcíny škodlivé účinky genetika MeSH
- otevřené čtecí rámce MeSH
- překrývající se geny * MeSH
- proteinové domény MeSH
- proteosyntéza MeSH
- vakcíny proti COVID-19 škodlivé účinky genetika MeSH
- virové geny * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Quinpirole (QNP) sensitization is a well-established model of stereotypical checking relevant to obsessive-compulsive disorder. Previously, we found that QNP-treated rats display deficits in hippocampus-dependent tasks. The present study explores the expression of immediate early genes (IEG) during QNP-induced stereotypical checking in the hippocampus, anterior cingulate cortex (ACC), orbitofrontal cortex (OFC), and medial prefrontal cortex (mPFC). Adult male rats were injected with QNP (0.5 mg/mL/kg; n = 15) or saline (n = 14) daily for 10 days and exposed to an arena enriched with two objects. Visits to the objects and the corners of the arena were recorded. QNP-treated rats developed an idiosyncratic pattern of visits that persisted across experimental days. On day 11, rats were exposed to the arena twice for 5 min and sacrificed. The expression of IEGs Arc and Homer1a was determined using cellular compartment analysis of temporal activity by fluorescence in situ hybridization. IEG-positive nuclei were counted in the CA1 area of the hippocampus, ACC, OFC, and mPFC. We found significantly fewer IEG-positive nuclei in the CA1 in QNP-treated rats compared to controls. The overlap between IEG expressing neurons was comparable between the groups. We did not observe significant differences in IEG expression between QNP treated and control rats in ACC, OFC, and mPFC. In conclusion, treatment of rats with quinpirole decreases plasticity-related activity in the hippocampus during stereotypical checking.
- MeSH
- antagonisté dopaminu D2 farmakologie MeSH
- chinpyrol farmakologie MeSH
- cingulární gyrus účinky léků fyziologie MeSH
- hipokampus účinky léků fyziologie MeSH
- neurony účinky léků metabolismus MeSH
- neuroplasticita účinky léků fyziologie MeSH
- okamžité časné geny MeSH
- pohybová aktivita účinky léků MeSH
- potkani Long-Evans MeSH
- prefrontální mozková kůra účinky léků fyziologie MeSH
- receptory dopaminu D2 metabolismus MeSH
- receptory dopaminu D3 antagonisté a inhibitory MeSH
- regulace genové exprese účinky léků MeSH
- stereotypní chování účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Bacteriophages of the significant veterinary pathogen Staphylococcus pseudintermedius are rarely described morphologically and genomically in detail, and mostly include phages of the Siphoviridae family. There is currently no taxonomical classification for phages of this bacterial species. Here we describe a new phage designated vB_SpsS_QT1, which is related to phage 2638A originally described as a Staphylococcus aureus phage. Propagating strain S. aureus 2854 of the latter was reclassified by rpoB gene sequencing as S. pseudintermedius 2854 in this work. Both phages have a narrow but different host range determined on 54 strains. Morphologically, both of them belong to the family Siphoviridae, share the B1 morphotype, and differ from other staphylococcal phage genera by a single long fibre at the terminus of the tail. The complete genome of phage vB_SpsS_QT1 was sequenced with the IonTorrent platform and expertly annotated. Its linear genome with cohesive ends is 43,029 bp long and encodes 60 predicted genes with the typical modular structure of staphylococcal siphophages. A global alignment found the genomes of vB_SpsS_QT1 and 2638A to share 84% nucleotide identity, but they have no significant similarity of nucleotide sequences with other phage genomes available in public databases. Based on the morphological, phylogenetic, and genomic analyses, a novel genus Fibralongavirus in the family Siphoviridae is described with phage species vB_SpsS_QT1 and 2638A.
The Deltaretrovirus genus of retroviruses (family Retroviridae) includes the human T cell leukemia viruses and bovine leukemia virus (BLV). Relatively little is known about the biology and evolution of these viruses, because only a few species have been identified and the genomic 'fossil record' is relatively sparse. Here, we report the discovery of multiple novel endogenous retroviruses (ERVs) derived from ancestral deltaretroviruses. These sequences-two of which contain complete or near complete internal coding regions-reside in genomes of several distinct mammalian orders, including bats, carnivores, cetaceans, and insectivores. We demonstrate that two of these ERVs contain unambiguous homologs of the tax gene, indicating that complex gene regulation has ancient origins within the Deltaretrovirus genus. ERVs demonstrate that the host range of the deltaretrovirus genus is much more extensive than suggested by the relatively small number of exogenous deltaretroviruses described so far, and allow the evolutionary timeline of deltaretrovirus-mammal interaction to be more accurately calibrated.
- MeSH
- Deltaretrovirus genetika MeSH
- endogenní retroviry genetika izolace a purifikace MeSH
- fylogeneze MeSH
- genom virový MeSH
- geny pX MeSH
- hostitelská specificita * MeSH
- lidé MeSH
- molekulární evoluce * MeSH
- paleontologie MeSH
- savci virologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
During 2008-2018, we collected samples from rodents and patients throughout the Czech Republic and characterized hantavirus isolates. We detected Dobrava-Belgrade and Puumala orthohantaviruses in patients and Dobrava-Belgrade, Tula, and Seewis orthohantaviruses in rodents. Increased knowledge of eco-epidemiology of hantaviruses will improve awareness among physicians and better outcomes of patients.
- MeSH
- fylogeneze MeSH
- hantavirové infekce epidemiologie imunologie virologie MeSH
- Hantavirus genetika imunologie MeSH
- imunoglobulin G imunologie MeSH
- imunoglobulin M imunologie MeSH
- lidé MeSH
- molekulární epidemiologie * MeSH
- protilátky virové MeSH
- virové geny MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- dopisy MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
The genomic characteristics of human cytomegalovirus (HCMV) strains sequenced directly from clinical pathology samples were investigated, focusing on variation, multiple-strain infection, recombination, and gene loss. A total of 207 datasets generated in this and previous studies using target enrichment and high-throughput sequencing were analyzed, in the process enabling the determination of genome sequences for 91 strains. Key findings were that (i) it is important to monitor the quality of sequencing libraries in investigating variation; (ii) many recombinant strains have been transmitted during HCMV evolution, and some have apparently survived for thousands of years without further recombination; (iii) mutants with nonfunctional genes (pseudogenes) have been circulating and recombining for long periods and can cause congenital infection and resulting clinical sequelae; and (iv) intrahost variation in single-strain infections is much less than that in multiple-strain infections. Future population-based studies are likely to continue illuminating the evolution, epidemiology, and pathogenesis of HCMV.
- MeSH
- cytomegalovirové infekce virologie MeSH
- Cytomegalovirus genetika MeSH
- databáze nukleových kyselin MeSH
- datové soubory jako téma MeSH
- DNA virů genetika MeSH
- genetická variace MeSH
- genom virový * genetika MeSH
- genotyp MeSH
- lidé MeSH
- molekulární evoluce MeSH
- mutace MeSH
- rekombinace genetická * MeSH
- sekvence nukleotidů * MeSH
- sekvenční analýza DNA MeSH
- sekvenování celého genomu MeSH
- virové geny MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Recent studies have shown that insects harbor numerous viruses of various taxa and that viral infections are often latent without noticeable symptoms. The red firebug Pyrrhocoris apterus, a true flightless bug from the family Pyrrhocoridae, is widely used for physiological studies on insect metabolism, endocrinology, and digestion. While exploring the transcriptome of P. apterus salivary glands, a nearly complete genomic sequence of a novel RNA virus was reconstructed. The virus, provisionally named Pyrrhocoris apterus virus 1 (PaV1), possesses eight potential open reading frames (ORFs) encoding for an array of proteins, some of which are involved in virus replication while others ensure success of the virus in multiple ways, including evasion of the host immune response. In addition to the information obtained from sequence analyses, we documented virus transmission, virus-induced mortality, host response upon persistent PaV1 infection, virion morphology, and putative virus-induced structures in salivary gland cells in a laboratory culture of red firebug. We propose that PaV1 belongs to a novel viral species of a new, yet-to-be established family.
Bovine leukemia virus (BLV) is a member of the genus Deltaretrovirus of the family Retroviridae and cause a chronic lymphosarcoma, which is extensive in cattle. In yaks (Bos grunniens), the distribution, strains and genetic characteristics of BLV have rarely been studied. The aim of our study was to investigate BLV infections in domestic yaks and determine the genetic variability of BLV circulating in a region of the Qinghai Tibet Plateau, China. Blood samples were collected from 798 yaks, which were from different farms from Gansu, Qinghai and Sichuan provinces surrounding the Qinghai-Tibet Plateau. Nested PCR targeting BLV long terminal repeats was used to detect the BLV provirus. The highest prevalence of BLV infection was in Gansu province, where it was 18.93% (39/206) in white yaks from Tianzhu City and 19.14% (31/162) in black yaks from Gannan City. In Qinghai and Sichuan provinces, the prevalence of BLV in black yaks was 14.83% (35/236) and 14.94% (29/194), respectively. The prevalence of BLV was not significantly different in yaks up to one year old than in older animals. Phylogenetic analysis was performed using 16 different env-gp51 (497-bp) gene sequences from the three provinces and 71 known BLV strains, which revealed that in both Gansu and Qinghai provinces, genotypes 6 and 10 of the BLV strains were at high levels, whereas only genotype 10 was prevalent in Sichuan Province. Phylogenetic analysis and sequence comparisons revealed 95.7-99.8% sequence identity among the full-length env genes of 16 strains, nearly full-length genome sequences of six BLV strains, and those of the known genotypes 6 and 10 of BLV. This study provides comprehensive information is regarding the widespread infection of domestic yaks with BLV on the Qinghai-Tibet Plateau of China, and shows that at least two BLV genotypes (genotypes 6 and 10) are circulating in this population.
- MeSH
- enzootická bovinní leukóza epidemiologie přenos virologie MeSH
- exprese genu MeSH
- fylogeneze * MeSH
- genotyp * MeSH
- geny env * MeSH
- koncové repetice MeSH
- molekulární epidemiologie MeSH
- prevalence MeSH
- sekvence nukleotidů MeSH
- sekvenční homologie nukleových kyselin MeSH
- sekvenční seřazení MeSH
- skot MeSH
- virus bovinní leukemie klasifikace genetika izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Tibet epidemiologie MeSH
Avian leukosis viruses (ALVs), which are pathogens of concern in domestic poultry, utilize specific receptor proteins for cell entry that are both necessary and sufficient for host susceptibility to a given ALV subgroup. This unequivocal relationship offers receptors as suitable targets of selection and biotechnological manipulation with the aim of obtaining virus-resistant poultry. This approach is further supported by the existence of natural knock-outs of receptor genes that segregate in inbred lines of chickens. We used CRISPR/Cas9 genome editing tools to introduce frame-shifting indel mutations into tva, tvc, and tvj loci encoding receptors for the A, C, and J ALV subgroups, respectively. For all three loci, the homozygous frame-shifting indels generating premature stop codons induced phenotypes which were fully resistant to the virus of respective subgroup. In the tvj locus, we also obtained in-frame deletions corroborating the importance of W38 and the four amino-acids preceding it. We demonstrate that CRISPR/Cas9-mediated knock-out or the fine editing of ALV receptor genes might be the first step in the development of virus-resistant chickens.
- MeSH
- buněčné linie MeSH
- CRISPR-Cas systémy * MeSH
- editace genu * MeSH
- genetické techniky MeSH
- genetické vektory genetika MeSH
- guide RNA, Kinetoplastida MeSH
- kur domácí MeSH
- odolnost vůči nemocem genetika MeSH
- ptačí leukóza genetika virologie MeSH
- sekvence nukleotidů MeSH
- virové geny MeSH
- virové receptory genetika metabolismus MeSH
- virus ptačí leukózy fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Murine herpesvirus 68 (MHV-68) is a natural pathogen that infects murid rodents, which serves as hosts for Ixodes ricinus ticks. For the first time, MHV-68 was detected in immature I. ricinus ticks feeding on Lacerta viridis lizards trapped in Slovakia, which supports the idea that ticks can acquire the virus from feeding on infected hosts. The recent discovery of MHV-68 infection and MHV-68 M3 gene transcripts in Dermacentor reticulatus ticks collected in Slovakia also supports this suggestion. Here, for the first time, we report MHV-68 infection, which was detected by nested PCR, in I. ricinus adults collected from the vegetation, and the viral load in infected ticks was determined by quantitative PCR. The viral incidence in ticks was 38.1% (21/55), and the viral load varied from 1.5 × 103 to 2.85 × 104 genome copies per tick. These results suggest that the I. ricinus ticks became infected with MHV-68 from biting infected rodents; thus, I. ricinus ticks may play a role in the spread of this virus in nature.
- MeSH
- arachnida jako vektory virologie MeSH
- DNA virů genetika MeSH
- hlodavci MeSH
- klíště virologie MeSH
- Rhadinovirus izolace a purifikace fyziologie MeSH
- virová nálož MeSH
- virové geny genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Slovenská republika MeSH
