Article
Microbiology
Baylee Heiden, Elke Muhlberger, Christopher W. Lennon, Adam J. Hume
Summary: Inteins are polypeptides that can interrupt protein sequences and remove themselves through protein splicing. In this study, a fluorescent protein was inserted within an intein, creating a recombinant Ebola virus expressing the fluorescent protein. Multiple potential insertion sites were tested, and a site within the VP30 gene was identified as efficient for intein splicing in mammalian cells while preserving VP30 function. A virus containing the fluorescent protein was successfully rescued, demonstrating a new intein-based application for adding reporters to systems without the need for additional genes.
Article
Virology
Chengcheng Zhang, Guoying Zhang, Yu Zhang, Xiaojing Lin, Xiujuan Zhao, Qinghua Cui, Lijun Rong, Ruikun Du
Summary: In this study, a reporter influenza A/H3N2 virus (A/NY-HiBiT) derived from a clinical isolate was constructed by placing a minimized HiBiT tag to the viral nuclear-export protein. The HiBiT tag did not affect the viral genome balance, and the recombinant A/NY-HiBiT virus remained stable. The replication profile of the HiBiT-tagged virus could be measured using a simple Nano-Glo assay, providing a robust platform for high-throughput screening. Using this platform, three fractions from Chinese medicinal materials were identified as potent anti-influenza A virus actives.
JOURNAL OF MEDICAL VIROLOGY
(2023)
Article
Immunology
Lochlain Corliss, Madeline Holliday, Nicholas J. J. Lennemann
Summary: Flavivirus infection causes dramatic remodeling of the ER, with viral replication occurring in virus-induced vesicular invaginations in the ER membrane. A new plasmid-based reporter system has been developed to monitor flavivirus infection and observe virus-induced manipulation of single cells in real-time. The system requires viral protease cleavage to release an ER-anchored fluorescent protein infection reporter.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2022)
Article
Immunology
Minjin Kim, Yucheol Cheong, Jinhee Lee, Jongkwan Lim, Sanguine Byun, Yo Han Jang, Baik Lin Seong
Summary: This study demonstrated that caspase-triggered live attenuated influenza vaccine induced broad reactive antibody response and provided heterosubtypic protection against diverse influenza viruses in mice model. The protection relied on non-neutralizing antibodies-mediated ADCC activities and robust mucosal secretion of sIgA, which highlight the potential of this vaccine as a promising option for developing a universal influenza vaccine.
FRONTIERS IN IMMUNOLOGY
(2021)
Article
Biochemistry & Molecular Biology
Erica Russo, Fabrice Lemaitre, Beatrice Corre, Aleksandra S. Chikina, Francina Langa-Vives, Philippe Bousso
Summary: SPICE-Met is a method for profiling energy metabolism in single cells using flow cytometry or imaging. It can infer the dependence of energy metabolism on oxidative phosphorylation and glycolysis, analyze immune metabolism, and dissect the heterogeneity and plasticity of energy metabolism in single macrophages.
Article
Chemistry, Analytical
Ui Jin Lee, Yunkwang Oh, Oh Seok Kwon, Yong-Beom Shin, Moonil Kim
Summary: We developed a highly sensitive and specific bimolecular fluorescence complementation (BiFC)-based influenza A virus (IAV)-sensing system. The system utilizes a galactose/glucose-binding protein (GGBP) with a N-terminal large domain (YN1-172) and a C-terminal small domain (YC173-239) made up of enhanced yellow fluorescence protein (eYFP). The system demonstrated ultra-high sensitivity and selective responses for IAV detection, making it a simple and effective platform for IAV detection without the need for virus culture or RNA extraction processes.
Article
Infectious Diseases
Tongtong Sun, Yanna Guo, Lingcai Zhao, Menglu Fan, Nan Huang, Miao Tian, Qingzheng Liu, Jingjin Huang, Zhiyuan Liu, Yongzhen Zhao, Zhiwei Ji, Jihui Ping
Summary: The PB1 gene in human H3N2 influenza viruses underwent specific changes from 2002-2009 that remained stable through 2019. By testing the functions of these mutations in different virus strains, it was found that certain mutations increased viral replication capacity while others reduced polymerase activity. This study demonstrates how identified PB1 mutations contributed to the evolution of human influenza A (H3N2) viruses.
TRANSBOUNDARY AND EMERGING DISEASES
(2022)
Article
Virology
Aitor Nogales, Michael Schotsaert, Raveen Rathnasinghe, Marta L. DeDiego, Adolfo Garcia-Sastre, Luis Martinez-Sobrido
Summary: The influenza A virus (IAV) can infect various mammalian and avian species, and studies have developed replication-competent IAV expressing traceable reporter genes to better understand its biology and pathogenesis. These novel approaches provide valuable tools for developing new therapeutic strategies against IAV infections.
Article
Veterinary Sciences
Lucas M. Ferreri, Silvia Carnaccini, Valeria Olivera, Ariel Pereda, Daniela Rajao, Daniel R. Perez
Summary: This study evaluated the adaptability of an H4N2 influenza A virus from the South American lineage to chickens. After a few passages, the virus acquired five mutations, resulting in enhanced infectivity in ex vivo trachea explants but overall decreased infection in lung explants. Infection in 3-week-old chickens lasted longer and was detected in more tissues, indicating adaptation of the H4N2 influenza A virus to chickens.
FRONTIERS IN VETERINARY SCIENCE
(2023)
Article
Microbiology
Min Zhu, Jianqiao He, Hao Zeng, Pingping Wang, Yaohui Zhu, Fanyuan Sun, Xin Huang, Yi Xu, Chongqiang Huang, Jiancai Chen, Xinyi Guo, Huabo Zhou, Zuzhang Wei, Kang Ouyang, Weijian Huang, Ying Chen
Summary: Mouse-adapted variants of swine-origin EA H1N1 influenza virus exhibited enhanced virulence and replication capability due to adaptive mutations primarily in the RNP, HA, and NS genes. These mutations enhanced polymerase activity and contributed to increased virulence, highlighting the importance of mutations in the HA and NS genes.
VETERINARY MICROBIOLOGY
(2023)
Article
Biology
Dorottya Anna Simon, Andras Talas, Peter Istvan Kulcsar, Zsuzsanna Biczok, Sarah Laura Krausz, Gyorgy Varady, Ervin Welker
Summary: Prime editing is a gene engineering tool that allows editing of the genome by introducing insertions, deletions, and substitutions. However, the efficiency of prime editing has been low, typically achieving editing rates of only 10-30%. To address this issue, researchers have developed PEAR, a fluorescent tool that can sensitively detect prime editing activity, improving the accuracy and applicability of editing.
Article
Developmental Biology
Colin J. Dinsmore, Philippe Soriano
Summary: Understanding how cells activate intracellular signaling pathways in response to external signals is an important goal in cell and developmental biology. Live-cell signaling reporters have greatly contributed to our understanding of signaling dynamics. Translocation reporters have received attention due to their ease of use and fast response times. This study created two new mouse reporter lines to investigate the ERK1/2 and AKT signaling pathways.
DEVELOPMENTAL BIOLOGY
(2023)
Article
Multidisciplinary Sciences
Alejandra Bernardi, Dino Gobelli, Julia Serna, Paulina Nawrocka, Gabriel March-Rossello, Antonio Orduna, Piotr Kozlowski, Maria Simarro, Miguel A. de la Fuente
Summary: The study presents a novel assay to measure HR in living cells by integrating HR substrate at a specific locus to quantify HR events, which can further investigate the mechanisms controlling HR.
Article
Chemistry, Multidisciplinary
Hao Chang, Xiao Hu, Xiaomei Tang, Shiwei Tian, Yidan Li, Xing Lv, Luqing Shang
Summary: In this study, a fluorescent probe called NAFP4 was developed to detect NADPH and visualize mitochondrial NADPH level changes in living cells. The study revealed that influenza virus infection leads to an increase in NADPH production in host cells, which is caused by an elevated level of G6PDH. Moreover, there was a positive correlation between G6PDH level and virus replication. This study highlights the influence of virus infection on host metabolism in NADPH production and suggests G6PDH as a potential target for antiviral therapy.
Article
Biochemical Research Methods
Anna Yudenko, Anastasia Smolentseva, Ivan Maslov, Oleg Semenov, Ivan M. Goncharov, Vera V. Nazarenko, Nina L. Maliar, Valentin Borshchevskiy, Valentin Gordeliy, Alina Remeeva, Ivan Gushchin
Summary: Protein-fragment complementation assays are commonly used to study protein-protein interactions. The development of CagFbFP, a small thermostable FbFP based on a LOV domain-containing protein, provides a split fluorescent reporter that can be used to investigate protein-protein interactions in anaerobic conditions without the need for exogenous fluorophores. This advancement in fluorescent protein technology may lead to further development of LOV and PAS domain-based fluorescent reporters and optogenetic tools.
ACS SYNTHETIC BIOLOGY
(2021)