Gene Editing of the Catfish Gonadotropin-Releasing Hormone Gene and Hormone Therapy to Control the Reproduction in Channel Catfish, <i>Ictalurus punctatus</i> by Guyu Qin, Zhenkui Qin, Cuiyu Lu, Zhi Ye, Ahmed Elaswad, Max Bangs, Hanbo Li, Yiliu Zhang, Yingqi Huang, Huitong Shi, Kamal Gosh, Nermeen Y. Abass, Khoi Vo, Ramjie Odin, William S. Bugg, Nathan J. C. Backenstose, David Drescher, Zachary Taylor, Timothy Braden, Baofeng Su, Rex A. Dunham

“…Targeted cfGnRH fish were sequenced and base deletion, substitution, and insertion were detected. The gene mutagenesis was achieved in 52.9% of P₁ fish. …”
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Genomic Analysis of a Novel Phage Infecting the Turkey Pathogen <i>Escherichia coli</i> APEC O78 and Its Endolysin Activity by Sangsang Deng, Qiang Xu, Yajuan Fu, Leiqin Liang, Yan Wu, Fang Peng, Meiying Gao

“…The results of site-directed mutagenesis indicated that LysO78 contained the catalytic residues E54 and E64. …”
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Identification of a Common Conformational Epitope on the Glycoprotein E2 of Classical Swine Fever Virus and Border Disease Virus by Yu-Liang Huang, Denise Meyer, Alexander Postel, Kuo-Jung Tsai, Hsin-Meng Liu, Chia-Huei Yang, Yu-Chun Huang, Nicholas Berkley, Ming-Chung Deng, Fun-In Wang, Paul Becher, Helen Crooke, Chia-Yi Chang

“…One CSFV-specific conformational epitope and one CSFV and BDV common epitope within domain B/C of E2 were identified. Site-directed mutagenesis confirmed that residues G725 and V738/I738 of the CSFV-specific epitope and P709/L709 and E713 of the second epitope are important for mAbs binding. …”
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Substrate‐enzyme interactions and catalytic mechanism in a novel family VI esterase with dibutyl phthalate-hydrolyzing activity by Jiliang Cheng, Huan Du, Meng-Sha Zhou, Yuan Ji, You-Qun Xie, He-Biao Huang, Shu-Hui Zhang, Fen Li, Lei Xiang, Quan-Ying Cai, Yan-Wen Li, Hui Li, Meng Li, Hai-Ming Zhao, Ce-Hui Mo

“…Based on the information on exact binding sites of DBP on PS06828 provided by molecular docking, the catalytic mechanism mediated by key residues (Ser113, Asp166, and His197) was proposed and subsequently confirmed by site-directed mutagenesis. The results can strengthen our mechanistic understanding of family VI esterase involved in hydrolysis of phthalic acid esters, and provide a solid foundation for further enzymatic modification.…”
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Cytotoxicity of Extracts from <i>Petiveria alliacea</i> Leaves on Yeast by Bruna B. F. Cal, Luana B. N. Araújo, Brenno M. Nunes, Claudia R. da Silva, Marcia B. N. Oliveira, Bianka O. Soares, Alvaro A. C. Leitão, Marcelo de Pádula, Debora Nascimento, Douglas S. A. Chaves, Rachel F. Gagliardi, Flavio J. S. Dantas

“…Three experimental assays were performed: cellular inactivation, mutagenesis (canavanine resistance system) and loss of mitochondrial function (petites colonies). …”
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Tumour mutations in long noncoding RNAs enhance cell fitness by Roberta Esposito, Andrés Lanzós, Tina Uroda, Sunandini Ramnarayanan, Isabel Büchi, Taisia Polidori, Hugo Guillen-Ramirez, Ante Mihaljevic, Bernard Mefi Merlin, Lia Mela, Eugenio Zoni, Lusine Hovhannisyan, Finn McCluggage, Matúš Medo, Giulia Basile, Dominik F. Meise, Sandra Zwyssig, Corina Wenger, Kyriakos Schwarz, Adrienne Vancura, Núria Bosch-Guiteras, Álvaro Andrades, Ai Ming Tham, Michaela Roemmele, Pedro P. Medina, Adrian F. Ochsenbein, Carsten Riether, Marianna Kruithof-de Julio, Yitzhak Zimmer, Michaela Medová, Deborah Stroka, Archa Fox, Rory Johnson

“…To directly evaluate the functional significance of NEAT1 SNVs, we use in cellulo mutagenesis to introduce tumour-like mutations in the gene and observe a significant and reproducible increase in cell fitness, both in vitro and in a mouse model. …”
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A Unique Mode of Coenzyme A Binding to the Nucleotide Binding Pocket of Human Metastasis Suppressor NME1 by Maria-Armineh Tossounian, Stefan Denchev Hristov, Jonathan Alexis Semelak, Bess Yi Kun Yu, Maria Baczynska, Yuhan Zhao, Dario Ariel Estrin, Madia Trujillo, Valeriy Filonenko, Jerome Gouge, Ivan Gout

“…Crystallographic studies suggested the involvement of arginine 58 and threonine 94 in mediating specific interactions with CoA. Site-directed mutagenesis and CoA-based affinity purifications showed that arginine 58 mutation to glutamate (R58E) and threonine 94 mutation to aspartate (T94D) prevent hNME1 from binding to CoA. …”
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Global Down-regulation of Gene Expression Induced by Mouse Mammary Tumor Virus (MMTV) in Normal Mammary Epithelial Cells by Waqar Ahmad, Neena G. Panicker, Shaima Akhlaq, Bushra Gull, Jasmin Baby, Thanumol A. Khader, Tahir A. Rizvi, Farah Mustafa

“…Comparison of our results with the Wnt1 mouse model revealed further insights into how MMTV expression could lead to activation of the Wnt1 pathway independent of insertional mutagenesis. Thus, the key pathways, DEGs, and hub genes identified in this study can provide important clues to elucidate the molecular mechanisms involved in MMTV replication, escape from cellular anti-viral response, and potential to cause cell transformation. …”
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The Endophyte Pantoea alhagi NX-11 Alleviates Salt Stress Damage to Rice Seedlings by Secreting Exopolysaccharides by Liang Sun, Liang Sun, Liang Sun, Peng Lei, Peng Lei, Peng Lei, Qian Wang, Qian Wang, Junjie Ma, Junjie Ma, Yijing Zhan, Yijing Zhan, Yijing Zhan, Kang Jiang, Kang Jiang, Kang Jiang, Zongqi Xu, Zongqi Xu, Zongqi Xu, Hong Xu, Hong Xu, Hong Xu

“…Interestingly, rice seedlings that were inoculated with an EPS-deficient strain named NX-11eps– that was obtained by atmospheric and room temperature plasma (ARTP) mutagenesis were damaged by salt stress and had similar physiological and biochemical indicators to the control group. …”
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PHÂN TÍCH DI TRUYỀN KHẢ NĂNG DI CHUYỂN THEO PHENANTHRENE CỦA VI KHUẨN PSEUDOMONAS STUTZERI E1 by Nguyễn Thị Phi Oanh, Dirk Springael, Line Nielsen, Paulina Estrada De Los Santos, René De Mot

“…A mutant bank of strain E1 was constructed using the pTnMod-OGm plasposon mutagenesis system.  82 out of 2639 electroporants screened showed an altered phenotype of phenanthrene-driven motility and/or phenanthrene degradation. 16 representative mutants were selected for sequencing the genes flanking the plasposon insertion. …”
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Cellulase hyper-production by Trichoderma reesei mutant SEU-7 on lactose by Chengcheng Li, Fengming Lin, Le Zhou, Lei Qin, Bingzhi Li, Zhihua Zhou, Mingjie Jin, Zhan Chen

“…Results Trichoderma reesei mutant SEU-7 was constructed from T. reesei RUT-C30 with the overexpression of endogenous gene β-glucosidase (BGL1) by insertional mutagenesis via Agrobacterium tumefaciens-mediated transformation (AMT). …”
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Identification of neutralizing epitopes on the D/A domain of the E2 glycoprotein of classical swine fever virus by Yu-Liang Huang, Denise Meyer, Alexander Postel, Kuo-Jung Tsai, Hsin-Meng Liu, Chia-Huei Yang, Yu-Chun Huang, Hui-Wen Chang, Ming-Chung Deng, Fun-In Wang, Paul Becher, Helen Crooke, Chia-Yi Chang

“…Three CSFV-specific epitopes, i.e., 780-IEEMGDDFGFGLCPF-794, 810-NGSAFYLVCPIGWTG-824, and 846-REKPF-850, were identified within the D/A domain of E2. Site-directed mutagenesis further confirmed that residues 783-MGD-785, 789-FGLCPF-794, 813-AFYLVCPIGWTG-824, and 846-REK-848 were critical residues in these regions. …”
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Pesticin-Like Effector VgrG3cp Targeting Peptidoglycan Delivered by the Type VI Secretion System Contributes to Vibrio cholerae Interbacterial Competition by Ming Liu, Meng-Yu Zhao, Heng Wang, Zeng-Hang Wang, Zhao Wang, Ying Liu, Yin-Peng Li, Tao Dong, Yang Fu

“…Using heterologous expression, protein secretion, and peptidoglycan-degrading assays, we demonstrated that VgrG3cp is a T6SS-dependent effector harboring cell wall muramidase activity and that its toxicity can be neutralized by cognate immunity protein TsiV3cp. Site-directed mutagenesis proved that the aspartic acid residue at position 867 is crucial for VgrG3cp-mediated antibacterial activity. …”
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Structure-guided glyco-engineering of ACE2 for improved potency as soluble SARS-CoV-2 decoy receptor by Tümay Capraz, Nikolaus F Kienzl, Elisabeth Laurent, Jan W Perthold, Esther Föderl-Höbenreich, Clemens Grünwald-Gruber, Daniel Maresch, Vanessa Monteil, Janine Niederhöfer, Gerald Wirnsberger, Ali Mirazimi, Kurt Zatloukal, Lukas Mach, Josef M Penninger, Chris Oostenbrink, Johannes Stadlmann

“…Engineering of ACE2 N-glycosylation by site-directed mutagenesis or glycosidase treatment resulted in enhanced binding affinities and improved virus neutralization without notable deleterious effects on the structural stability and catalytic activity of the protein. …”
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Ribavirin inhibits the replication of infectious bursal disease virus predominantly through depletion of cellular guanosine pool by Towseef Akram, Irfan Gul, Irfan Gul, Mahrukh Parveez Zia, Mahrukh Parveez Zia, Amreena Hassan, Amreena Hassan, Amina Khatun, Riaz Ahmad Shah, Syed Mudasir Ahmad, Nazir Ahmad Ganai, Naveed Anjum Chikan, Won-Il Kim, Nadeem Shabir

“…Ribavirin inhibits ds- RNA viruses, likely through IMPDH inhibition and not mutagenesis. The inhibitory effect may, however, be augmented by other non-mutagenic mechanisms, like induction of antiviral cytokines in chicken embryo fibroblasts or interaction of ribavirin with the active sites of RNA-dependent RNA polymerase of the virus.…”
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TYK2 Variants in B-Acute Lymphoblastic Leukaemia by Edgar Turrubiartes-Martínez, Irene Bodega-Mayor, Pablo Delgado-Wicke, Francisca Molina-Jiménez, Diana Casique-Aguirre, Martín González-Andrade, Inmaculada Rapado, Mireia Camós, Cristina Díaz-de-Heredia, Eva Barragán, Manuel Ramírez-Orellana, Beatriz Aguado, Ángela Figuera, Joaquín Martínez-López, Elena Fernández-Ruiz

“…To functionally characterise them, they were generated by direct mutagenesis, cloned in expression vectors, and transfected in TYK2-deficient cells. …”
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Deciphering the Binding of Salicylic Acid to <i>Arabidopsis thaliana</i> Chloroplastic GAPDH-A1 by Igor Pokotylo, Denis Hellal, Tahar Bouceba, Miguel Hernandez-Martinez, Volodymyr Kravets, Luis Leitao, Christophe Espinasse, Isabelle Kleiner, Eric Ruelland

“…A simulation in water of the complex between SA and the protein allowed us to determine that only one pocket—a surface cavity around Asn35—would efficiently bind SA in the presence of solvent. In silico mutagenesis and simulations of the ligand/protein complexes pointed to the importance of Asn35 and Arg81 in the binding of SA to GAPA1. …”
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Structure-guided manipulation of the regioselectivity of the cyclosporine A hydroxylase CYP-sb21 from Sebekia benihana by Fengwei Li, Li Ma, Xingwang Zhang, Jingfei Chen, Feifei Qi, Yinyue Huang, Zepeng Qu, Lishan Yao, Wei Zhang, Eung-Soo Kim, Shengying Li

“…Together with sequence and 3D structure comparisons, Autodock-based substrate docking, molecular dynamics (MD) simulation, and site-directed mutagenesis, we identified a number of key residues including R294, E264, and M179 that can improve catalytic efficiency or change the regioselectivity of CYP-sb21 towards CsA, setting the stage for better enzymatic preparation of CsA-4-OH. …”
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Structure and physiological function of the human KCNQ1 channel voltage sensor intermediate state by Keenan C Taylor, Po Wei Kang, Panpan Hou, Nien-Du Yang, Georg Kuenze, Jarrod A Smith, Jingyi Shi, Hui Huang, Kelli McFarland White, Dungeng Peng, Alfred L George, Jens Meiler, Robert L McFeeters, Jianmin Cui, Charles R Sanders

“…Here, we report and validate the experimental three-dimensional structure of the human KCNQ1 voltage-gated potassium channel VSD in the intermediate state. We also used mutagenesis and electrophysiology in Xenopus laevisoocytes to functionally map the determinants of S4 helix motion during voltage-dependent transition from the intermediate to the activated state. …”
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Tyrosine 146 of the Human Na⁺/Taurocholate Cotransporting Polypeptide (NTCP) Is Essential for Its Hepatitis B Virus (HBV) Receptor Function and HBV Entry into Hepatocyte... by Dariusz Zakrzewicz, Regina Leidolf, Sebastian Kunz, Simon Franz Müller, Anita Neubauer, Silke Leiting, Nora Goldmann, Felix Lehmann, Dieter Glebe, Joachim Geyer

“…To study the role of Y139, Y141 and Y146 amino acids in NTCP biology, the aforementioned residues were substituted with alanine, phenylalanine or glutamate (mimicking phosphorylation) using site-directed mutagenesis. Similar to wt NTCP, the Y139A, Y141A, Y146A, Y141F, Y146F, and Y146E mutants were expressed at the plasma membrane of HEK293 cells and exhibited intact bile acid transport function. …”
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