A Review of Unreduced Gametes and Neopolyploids in Alfalfa: How to Fill the Gap between Well-Established Meiotic Mutants and Next-Generation Genomic Resources by Fabio Palumbo, Elisa Pasquali, Emidio Albertini, Gianni Barcaccia

“…Here, we propose a reverse genetics approach, by exploiting the genomic and transcriptomic resources available in alfalfa. …”
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Reovirus Type 3 Dearing Variants Do Not Induce Necroptosis in RIPK3-Expressing Human Tumor Cell Lines by Diana J. M. van den Wollenberg, Vera Kemp, Martijn J. W. E. Rabelink, Rob C. Hoeben

“…While this treatment induced necroptosis, three different reovirus T3D variants, i.e., the plasmid-based reverse genetics generated virus (T3D^K), the wild-type reovirus T3D isolate R124, and the junction adhesion molecule-A-independent reovirus mutant (jin-1) failed to induce necroptosis in HT29 cells. …”
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THE BIOSYNTHESIS REGULATION OF POTATO STEROIDAL GLYCOALKALOIDS by K. A. Ivanova, S. V. Gerasimova, E. K. Khlestkina

“…In the potato genome, 14 corresponding genes were identified, 8 of which were studied in detail using reverse genetics approaches. As a promising target for reducing SGA levels in tubers, the genes encoding PGA enzymes (belonging to the CYP72 subfamily cytochrome-P450-dependent monooxygenases catalyzing the conversion of hydrocholesterol to trihydrocholesterol) and SGT (SGA glycosyltransferases that catalyze the conversion of solanidine to its toxic glycosylated derivatives α-solanine and α-chaconine) are considered. …”
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Neuraminidase (NA) 370-Loop Mutations of the 2009 Pandemic H1N1 Viruses Affect NA Enzyme Activity, Hemagglutination Titer, Mouse Virulence, and Inactivated-Virus Immunogenicity by Ting-Hsuan Chen, Chung-Chu Chen, Suh-Chin Wu

“…In this study, we used PR8-based reverse genetics (RG) by replacing the HA and NA genes of A/Texas/05/2009 pH1N1 virus to obtain the wild-type pH1N1 and three NA 370-loop mutant viruses of pH1N1 (I365T/S366N), RG pH1N1 (I365E/S366D), and RG pH1N1 (I365T/S366A). …”
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Identification of the Genome Segments of Bluetongue Virus Type 26/Type 1 Reassortants Influencing Horizontal Transmission in a Mouse Model by Houssam Attoui, Baptiste Monsion, Bernard Klonjkowski, Stéphan Zientara, Peter P. C. Mertens, Fauziah Mohd Jaafar

“…These atypical viruses are transmitted horizontally by direct contact between infected and susceptible hosts (primarily small ruminants) causing only mild clinical signs, although the exact transmission mechanisms involved have yet to be determined. We used reverse genetics to generate a strain of BTV-1 (BTV-1 RG_C7) which is less virulent, infecting IFNAR^(−/−) mice without killing them. …”
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NS1 and PA-X of H1N1/09 influenza virus act in a concerted manner to manipulate the innate immune response of porcine respiratory epithelial cells by Robin Avanthay, Robin Avanthay, Robin Avanthay, Obdulio Garcia-Nicolas, Obdulio Garcia-Nicolas, Gert Zimmer, Gert Zimmer, Artur Summerfield, Artur Summerfield

“…In this study, we took advantage of reverse genetics to generate three LAIV based on the pandemic H1N1 2009 (pH1N1/09) virus strain: ΔPA-X, which is defective in the synthesis of the accessory PA-X protein, NS1(1-126) lacking 93 amino acids at the C-terminus of the NS1 protein, and a combination of both. …”
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Effect of salinity stress in Setaria viridis (L.) P. Beauv. accession A10.1 during seed germination and plant development by Thalita Massaro Malheiros Ferreira, Mariana de Lima Santos, Cecilia Lima Lopes, Carlos Antonio Ferreira de Sousa, Manoel Teixeira Souza Junior

“…Beauv. is a species proposed to be used as model plant in reverse genetics studies for the validation of gene function. …”
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CRISPR-based tools for targeted genetic manipulation in pathogenic Sporothrix species by Remi Hatinguais, Ian Leaves, Gordon D. Brown, Alistair J. P. Brown, Matthias Brock, Roberta Peres da Silva

“…Although certain virulence factors have been proposed as potential determinants for sporotrichosis, the scarcity of molecular tools for performing reverse genetics in Sporothrix has significantly impeded the dissection of mechanisms underlying the disease. …”
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Screening of Porcine Innate Immune Adaptor Signaling Revealed Several Anti-PRRSV Signaling Pathways by Yulin Xu, Mengxue Ye, Youwen Zhang, Shaohua Sun, Jia Luo, Sen Jiang, Jiajia Zhang, Xueliang Liu, Qi Shao, Qi Cao, Wanglong Zheng, François Meurens, Nanhua Chen, Jianzhong Zhu

“…To better quantify the PRRSV replication, the GFP signal of PRRSV from reverse genetics were measured by flow cytometry and similarly varying anti-PRRSV activities by different signaling adaptors were observed. …”
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Evolution and Application of Genome Editing Techniques for Achieving Food and Nutritional Security by Sajid Fiaz, Sunny Ahmar, Sajjad Saeed, Aamir Riaz, Freddy Mora-Poblete, Ki-Hung Jung

“…Future development and the optimization of forward and reverse genetics to achieve food security are evaluated.…”
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Experimental Adaptation of Murine Norovirus to Calcium Hydroxide by Wakana Oishi, Mikiko Sato, Kengo Kubota, Kengo Kubota, Ryoka Ishiyama, Reiko Takai-Todaka, Kei Haga, Kazuhiko Katayama, Daisuke Sano, Daisuke Sano, Daisuke Sano

“…The infectious clones with the K345R produced using a plasmid-based reverse genetics system exhibited greater tolerance in a lime solution, which indicated that the specific amino acid substitution was solely involved in the viral tolerance in lime treatment.…”
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Construction, Characterization and Application of Recombinant Porcine Deltacoronavirus Expressing Nanoluciferase by Puxian Fang, Huichang Zhang, He Sun, Gang Wang, Sijin Xia, Jie Ren, Jiansong Zhang, Liyuan Tian, Liurong Fang, Shaobo Xiao

“…Here, we successfully generated an infectious clone of PDCoV strain CHN-HN-2014 using a combination of bacterial artificial chromosome (BAC)-based reverse genetics system with a one-step homologous recombination. …”
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Tissue-Nonspecific Alkaline Phosphatase, a Possible Mediator of Cell Maturation: Towards a New Paradigm by Masahiro Sato, Issei Saitoh, Yuki Kiyokawa, Yoko Iwase, Naoko Kubota, Natsumi Ibano, Hirofumi Noguchi, Youichi Yamasaki, Emi Inada

“…Mutations in the gene coding for TNSALP are thought to be responsible for the abnormalities, suggesting the essential role of TNSALP in these events. Moreover, a reverse-genetics-based study using mice revealed that TNSALP is important in bone and tooth development as well as neurogenesis. …”
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Genetic manipulation of porcine deltacoronavirus reveals insights into NS6 and NS7 functions: a novel strategy for vaccine design by Mengjia Zhang, Wan Li, Peng Zhou, Dejian Liu, Rui Luo, Anan Jongkaewwattana, Qigai He

“…Taken together, the reverse genetics platform described here not only provides more insights into the role of PDCoV accessory proteins in viral replication and pathogenesis, but also allows the development of novel vaccines against PDCoV infection.…”
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Improved cold tolerance in switchgrass by a novel CCCH-type zinc finger transcription factor gene, PvC3H72, associated with ICE1–CBF–COR regulon and ABA-responsive genes by Zheni Xie, Wenjing Lin, Guohui Yu, Qiang Cheng, Bin Xu, Bingru Huang

“…The objectives of this study were to characterize PvC3H72 using reverse genetics approach and to understand its functional role in cold signal transduction and cold tolerance in switchgrass. …”
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Random Mutagenesis as a Promising Tool for Microalgal Strain Improvement towards Industrial Production by Mafalda Trovão, Lisa M. Schüler, Adriana Machado, Gabriel Bombo, Sofia Navalho, Ana Barros, Hugo Pereira, Joana Silva, Filomena Freitas, João Varela

“…While classical methods of forward genetics have been extensively used to determine gene function of randomly generated mutants, reverse genetics has been explored to generate specific mutations and target phenotypes. …”
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Steps of the Replication Cycle of the Viral Haemorrhagic Septicaemia Virus (VHSV) Affecting Its Virulence on Fish by Carmen López-Vázquez, Isabel Bandín, Valentina Panzarin, Anna Toffan, Argelia Cuenca, Niels J. Olesen, Carlos P. Dopazo

“…Many studies have been carried out to determine the participation of each gene in the VHSV virulence, most of them based on genome sequence analysis and/or reverse genetics to construct specific mutants and to evaluate their virulence phenotype. …”
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CRISPR-Cas-Guided Mutagenesis of Chromosome and Virulence Plasmid in Shigella flexneri by Cytosine Base Editing by Atin Sharma, Ruqiya Omer Aden, Andrea Puhar, David A. Cisneros

“…Much of the information about the genes dictating the pathophysiology of Shigella, both on the chromosome and the virulence plasmid, was obtained by classical reverse genetics. However, technical limitations of the prevalent mutagenesis techniques restrict the generation of mutants in a single reaction to a small number, preventing large-scale targeted mutagenesis of Shigella and the subsequent assessment of phenotype. …”
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Temperature Sensitivity: A Potential Method for the Generation of Vaccines against the Avian Coronavirus Infectious Bronchitis Virus by Sarah Keep, Phoebe Stevenson-Leggett, Angela Steyn, Michael S. Oade, Isobel Webb, Jamie Stuart, Lonneke Vervelde, Paul Britton, Helena J. Maier, Erica Bickerton

“…The <i>Gammacoronavirus</i> infectious bronchitis virus (IBV) is a highly contagious economically important respiratory pathogen of domestic fowl. Reverse genetics allows for the molecular study of pathogenic determinants to enable rational vaccine design. …”
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A C. albicans TRAPP Complex-Associated Gene Contributes to Cell Wall Integrity, Hyphal and Biofilm Formation, and Tissue Invasion by Dakota Ma, Miranda Yu, Susan Eszterhas, Christiane Rollenhagen, Samuel A. Lee

“…In S. cerevisiae, TCA17 encodes a transport protein particle (TRAPP) complex-associated protein. Using a reverse genetics approach with CRISPR-Cas9-mediated gene deletion, we analyzed the function of the TCA17 homolog in C. albicans. …”
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