An Engineered Influenza a Virus Expressing the Co-Stimulator OX40L as an Oncolytic Agent Against Hepatocellular Carcinoma by Yang H, Lei G, Deng Z, Sun F, Tian Y, Cheng J, Yu H, Li C, Bai C, Zhang S, An G, Yang P

“…Moreover, an increasing number of studies have shown that the therapeutic efficacy of OV can be further improved by arming OVs with immune-stimulating molecules.Methods: In this study, we used reverse genetics to produce a novel influenza A virus, termed IAV-OX40L, which contained the immune-stimulating molecule OX40L gene in the influenza virus nonstructural (NS1) protein gene. …”
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Metallothionein1A Regulates Rhizobial Infection and Nodulation in <i>Phaseolus vulgaris</i> by Citlali Fonseca-García, Claudia Marina López-García, Ronal Pacheco, Elisabeth Armada, Noreide Nava, Rocío Pérez-Aguilar, Jorge Solis-Miranda, Carmen Quinto

“…Here, we analyzed the role of the <i>PvMT1A</i> gene in <i>Phaseolus vulgaris</i>-<i>Rhizobium tropici</i> symbiosis using bioinformatics and reverse genetics approaches. Using in silico analysis, we identified six genes encoding MTs in <i>P. vulgaris</i>, which were clustered into three of the four classes described in plants. …”
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Systems-Wide Dissection of Organic Acid Assimilation in Pseudomonas aeruginosa Reveals a Novel Path To Underground Metabolism by Stephen K. Dolan, Andre Wijaya, Michael Kohlstedt, Lars Gläser, Paul Brear, Rafael Silva-Rocha, Christoph Wittmann, Martin Welch

“…In this work, we used a combination of ‘omics approaches (transcriptomics, proteomics, metabolomics, and 13C-fluxomics) and reverse genetics to provide systems-level insight into how the infection-relevant organic acids succinate and propionate are metabolized by Pa. …”
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A Tomato EMS-Mutagenized Population Provides New Valuable Resources for Gene Discovery and Breeding of Developmental Traits by Rocío Fonseca, Carmen Capel, Roberto Nieto-Canseco, Ana Ortiz-Atienza, Sandra Bretones, Juan D. López-Fábregas, Abraham S. Quevedo-Colmena, Ricardo Lebrón, Teresa Barragán-Lozano, Víctor Villalobos-Ramírez, Fernando J. Yuste-Lisbona, Trinidad Angosto, Juan Capel, Rafael Lozano

“…This lack of functional characterization hampers the use of the huge amount of genomic information available to improve the quality and productivity of this crop. Reverse genetics strategies such as artificial mutagenesis and next-generation sequencing approaches build the perfect tandem for increasing knowledge on functional annotation of tomato genes. …”
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Assembly-defective Tembusu virus ectopically expressing capsid protein is an approach for live-attenuated flavivirus vaccine development by Yu He, Jiaqi Guo, Xiaoli Wang, Senzhao Zhang, Li Mao, Tao Hu, Mingshu Wang, Renyong Jia, Dekang Zhu, Mafeng Liu, Xinxin Zhao, Qiao Yang, Ying Wu, Shaqiu Zhang, Juan Huang, Sai Mao, Xumin Ou, Qun Gao, Di Sun, Anchun Cheng, Shun Chen

“…In the present study, we demonstrated a method for generating flavivirus LAVs based on breaking spatially and temporally regulated C-prM cleavage to disturb the viral assembly process, using an avian flavivirus (Tembusu virus) as the model. Using reverse genetics technology, we successfully generated two recombinant viruses (CQW1-IRES-mC and CQW1-MINI-mC) with bicistronic genomic RNA in which native capsid genes were deleted and instead expressed in the 3’UTR under the control of an internal ribosome entry site (IRES) or minimum IRES. …”
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Rice grain yield and quality improvement via CRISPR/Cas9 system: an updated review by Aqib ZEB, Shakeel AHMAD, Javaria TABBASUM, Zhonghua SHENG, Peisong HU

“…Genomic alteration employing genome editing technologies (GETs) like CRISPR/Cas9 for reverse genetics has opened new avenues in agricultural sciences such as rice yield and grain quality improvement. …”
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Identification of a novel binding inhibitor that blocks the interaction between hSCARB2 and VP1 of enterovirus 71 by Qi Tang, Zhichao Xu, Fan Zhang, Yang Cai, Yinuo Chen, Baojing Lu, Hai-bing Zhou, Ke Lan, Shuwen Wu

“…The mechanism of action of PTC-209HBr was confirmed by time-of-addition, resistance selection and reverse genetics experiments, microscale thermophoresis (MST), viral binding and entry assays, coimmunoprecipitation (Co-IP) and immunofluorescence experiments (IF). …”
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Effect of D222G Mutation in the Hemagglutinin Protein on Receptor Binding, Pathogenesis and Transmissibility of the 2009 Pandemic H1N1 Influenza Virus by Belser, Jessica A., Pappas, Claudia, Zeng, Hui, Cox, Nancy J., Katz, Jacqueline M., Tumpey, Terrence M., Jayaraman, Akila, Raman, Rahul, Sasisekharan, Ram

“…To assess the relative contribution of this substitution in virus pathogenesis, transmission, and tropism, we introduced D222G by reverse genetics in the wild-type HA of the 2009 H1N1 virus, A/California/04/09 (CA/04). …”
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Investigating TDP43 biological dysfunction through the characterisation of Tardbp ENU mouse mutants: implications for neurodegeneration by De Sao Jose Martinho De Oliveira, H, Hugo De Sao Jose Martinho De Oliveira

“…</p> <p>In this thesis a reverse genetics approach was taken and two <em>Tardbp</em> ENU mutant mouse models were characterised, <em>Tardbp^F210I</em> (mutation in the RRM2 of TDP43) and <em>Tardbp^M323K</em> (mutation in the C-terminal domain of TDP43). …”

Genetic and chemical validation of Plasmodium falciparum aminopeptidase PfA-M17 as a drug target in the hemoglobin digestion pathway by Rebecca CS Edgar, Ghizal Siddiqui, Katheryn Hjerrild, Tess R Malcolm, Natalie B Vinh, Chaille T Webb, Clare Holmes, Christopher A MacRaild, Hope C Chernih, Willy W Suen, Natalie A Counihan, Darren J Creek, Peter J Scammells, Sheena McGowan, Tania F de Koning-Ward

“…Here, we use both reverse genetics and a compound specifically designed to inhibit the activity of PfA-M17 to show that PfA-M17 is essential for P. falciparum survival as it provides parasites with free amino acids for growth, many of which are highly likely to originate from hemoglobin. …”
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Vesicular stomatitis virus-based vaccine targeting plasmodium blood-stage antigens elicits immune response and protects against malaria with protein booster strategy by Yifan Sun, Yifan Sun, Xiaodan Shi, Feng Lu, Haitian Fu, Haitian Fu, Yi Yin, Jiahui Xu, Cheng Jin, Eun-taek Han, Xuan Huang, Yongquan Chen, Chunsheng Dong, Yang Cheng

“…In this study, we have used the reverse genetics system to construct recombinant VSVs (rVSVs) expressing apical membrane protein 1 (AMA1), rhoptry neck protein 2 (RON2), and reticulocyte-binding protein homolog 5 (RH5), which are required for Plasmodium falciparum invasion. …”
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Structure-Based Screening of Plasmodium berghei Glutathione S-Transferase Identifies CB-27 as a Novel Antiplasmodial Compound by Emilee E. Colón-Lorenzo, Daisy D. Colón-López, Joel Vega-Rodríguez, Alice Dupin, David A. Fidock, David A. Fidock, Abel Baerga-Ortiz, José G. Ortiz, Jürgen Bosch, Jürgen Bosch, Jürgen Bosch, Adelfa E. Serrano

“…This study aims to better understand the biological function of Plasmodium GST, assess its potential as a drug target, and identify novel antiplasmodial compounds using the rodent model P. berghei. By using reverse genetics, we provided evidence that GST is essential for survival of P. berghei intra-erythrocytic stages and is a valid target for drug development. …”
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Minor envelope proteins from GP2a to GP4 contribute to the spread pattern and yield of type 2 PRRSV in MARC-145 cells by Yuan-Zhe Bai, Yuan-Zhe Bai, Yue Sun, Yong-Gang Liu, Hong-Liang Zhang, Tong-Qing An, Qian Wang, Zhi-Jun Tian, Xinyuan Qiao, Xue-Hui Cai, Xue-Hui Cai, Yan-Dong Tang, Yan-Dong Tang, Yan-Dong Tang

“…By replacing the viral envelope genes with a reverse genetics system, we discovered that the minor envelope proteins, from GP2a to GP4, play a crucial role in determining the spread pattern and yield of type 2 PRRSV in MARC-145 cells. …”
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p21-activated kinase is involved in the sporulation, pathogenicity, and stress response of Arthrobotrys oligospora under the indirect regulation of Rho GTPase-activating protein by Meichen Zhu, Meichen Zhu, Yankun Liu, Yankun Liu, Xuewei Yang, Xuewei Yang, Lirong Zhu, Lirong Zhu, Yanmei Shen, Yanmei Shen, Shipeng Duan, Shipeng Duan, Jinkui Yang, Jinkui Yang

“…Herein, we characterized two orthologous Rho-GAPs (AoRga1 and AoRga2) and two PAKs (AoPak1 and AoPak2) through bioinformatics analysis and reverse genetics in Arthrobotrys oligospora, a typical nematode-trapping (NT) fungus. …”
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An Expeditious Neutralization Assay for Porcine Reproductive and Respiratory Syndrome Virus Based on a Recombinant Virus Expressing Green Fluorescent Protein by Juan Wang, Jiecong Yan, Shuaiyong Wang, Ronglin Chen, Yanru Xing, Qingyan Liu, Shuolei Gao, Yuxiang Zhu, Jiannan Li, Yanjun Zhou, Tongling Shan, Wu Tong, Hao Zheng, Ning Kong, Yifeng Jiang, Changlong Liu, Guangzhi Tong, Hai Yu

“…Here, a PRRSV expressing eGFP was successfully rescued with reverse genetics based on the infectious clone HuN4-F112-eGFP which we constructed. …”
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A recombinant murine-like rotavirus with Nano-Luciferase expression reveals tissue tropism, replication dynamics, and virus transmission by Yinxing Zhu, Liliana Sánchez-Tacuba, Liliana Sánchez-Tacuba, Liliana Sánchez-Tacuba, Gaopeng Hou, Takahiro Kawagishi, Takahiro Kawagishi, Takahiro Kawagishi, Ningguo Feng, Ningguo Feng, Ningguo Feng, Harry B. Greenberg, Harry B. Greenberg, Harry B. Greenberg, Siyuan Ding

“…Here, we report the generation of the first recombinant murine-like RV that encodes a Nano-Luciferase reporter (NLuc) using a newly optimized RV reverse genetics system. The NLuc-expressing RV was replication-competent in cell culture and both infectious and virulent in neonatal mice in vivo. …”
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Pathogenicity, transmissibility, and immunogenicity of recombinant H9N2 avian influenza viruses based on representative viruses of Southeast China by Hongyang Liu, Yikai Chen, Hongyang Li, Lingen Yang, Shili Yang, Xiaoying Luo, Song Wang, Ji-Long Chen, Shihong Yan

“…In order to use reverse genetics to rapid preparation of seed strains for vaccine production, and intend to prevent and control the H9N2 subtype epidemic strains of avian influenza virus (AIV). …”
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Impact of Individual Viral Gene Segments from Influenza A/H5N8 Virus on the Protective Efficacy of Inactivated Subtype-Specific Influenza Vaccine by Yassmin Moatasim, Ahmed Kandeil, Ahmed Mostafa, Omnia Kutkat, Mohamed El Sayes, Ahmed N. El Taweel, Maha AlKhazindar, Elsayed T. AbdElSalam, Rabeh El-Shesheny, Ghazi Kayali, Mohamed A. Ali

“…The contribution of the other internal proteins from H5N8, either individually or in combination, to the overall protective efficacy of PR8-based H5N8 vaccine has not been investigated. Using reverse genetics, a set of PR8-based vaccines expressing the individual proteins from an H5N8 strain were rescued and compared to the parent PR8 and low pathogenic H5N8 strains and the commonly used PR8_HA/NA-H5N8. …”
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PRAT Proteins Operate in Organellar Protein Import and Export in <i>Arabidopsis thaliana</i> by Claudia Rossig, John Gray, Oscar Valdes, Armin Springer, Sachin Rustgi, Diter von Wettstein, Christiane Reinbothe, Joachim Rassow, Steffen Reinbothe

“…Using a combination of reverse genetics, protein biochemistry and physiological tools, evidence was obtained for a role of chloroplast HP20, HP30 and HP30-2 in protein, but not amino acid, import into chloroplasts. …”
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Selection of an Optimal Recombinant Egyptian H9N2 Avian Influenza Vaccine Strain for Poultry with High Antigenicity and Safety by Se-Hee An, Seung-Eun Son, Jin-Ha Song, Seung-Min Hong, Chung-Young Lee, Nak-Hyung Lee, Young-Ju Jeong, Jun-Gu Choi, Youn-Jeong Lee, Hyun-Mi Kang, Kang-Seuk Choi, Hyuk-Joon Kwon

“…For the development of an optimized Egyptian H9N2 vaccine candidate virus for poultry, various recombinant Egyptian H9N2 viruses generated by a PR8-based reverse genetics system were compared in terms of their productivity and biosafety since Egyptian H9N2 avian influenza viruses already possess mammalian pathogenicity-related mutations in the hemagglutinin (HA), neuraminidase (NA), and PB2 genes. …”
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