Article
Fisheries
Jonathan Molcho, Rivka Manor, Maayan Shamsian, Gurucharan Sudarshan, Rivka Ofir, Danit Parker, Simy Weil, Hanin Wattad, Emily Hayun, Tom Levy, Eliahu D. Aflalo, Ayal Hendel, Amir Sagi
Summary: This study successfully established two CRISPR platforms for the giant freshwater prawn Macrobrachium rosenbergii and achieved high gene editing efficiency through optimization. Editing efficiency was examined and reached as high as 100% in embryos, while the maximum efficiency obtained in primary embryonic cell cultures was 64%. Additionally, a striking difference in deletion patterns around the Cas9 cut site was observed between the two systems, indicating distinct repair mechanisms that require further clarification. The investigation of the Pax6 transcription factor provided a phenotypic proof of concept by showing clear effects on eye development in edited embryos and larvae. This study lays down the foundations for precise functional genomic research and genome editing applications in crustacean species for both aquaculture and sustainable biocontrol, offering opportunities for the creation of selected crustacean lines with distinct attributes.
Article
Nanoscience & Nanotechnology
Min Zhu, Xiuxiu Wang, Ruosen Xie, Yuyuan Wang, Xianghui Xu, Jacobus Burger, Shaoqin Gong
Summary: This study presents a lipopeptide-based nanoparticle (LNP) that can efficiently deliver CRISPR Cas9/sgRNA ribonucleoprotein (RNP) for genome editing both in vitro and in vivo.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Biochemical Research Methods
Duran Zhao, Johanna L. Jones, Robert J. Gasperini, Jac C. Charlesworth, Guei-Sheung Liu, Kathryn P. Burdon
Summary: Identification of genes causing cataract is crucial for understanding the pathology of inherited pediatric cataract and developing new therapies. This study presents a step-by-step pipeline to evaluate cataract candidate genes in zebrafish using CRISPR-Cas9, offering a simple and efficient method for assessment in basic laboratory settings.
Article
Multidisciplinary Sciences
Nahyun Lee, Jiyeun Park, Jung-Eun Kim, Ji Young Shin, Kyunghun Min, Hokyoung Son
Summary: This study describes a CRISPR-Cas9-based genome-editing tool for direct delivery of preassembled Cas9 ribonucleoproteins (RNPs) into the protoplasts of F. graminearum. The use of RNPs significantly improves the efficiency of gene replacement and allows for large-scale functional analysis and difficult gene deletion.
Article
Biochemistry & Molecular Biology
Marina Zoppo, Nicole Okoniewski, Stanislav Pantelyushin, Johannes vom Berg, Kristin Schirmer
Summary: This study developed a strategy to CRISPR/Cas9 gene edit rainbow trout cell lines, achieving a gene editing efficiency of 39%. By using cloning cylinders, two monoclonal CRISPR edited rainbow trout cell lines were established for the first time, confirming the disruption of the cyp1a1 gene.
CELL AND BIOSCIENCE
(2021)
Article
Genetics & Heredity
Parthasarathy Santhanam, Caroline Labbe, Luciano Gomes Fietto, Richard R. Belanger
Summary: The study investigated the role of flocculosin in the biocontrol activity of P. flocculosa using two different approaches. Results showed that editing fat1 gene by HR led to abnormal phenotypes and inability to produce sporidia, while CRISPR-Cas9 transformation specifically affected flocculosin synthesis. The findings suggest that flocculosin-mediated antibiosis is not responsible for the mode of action of P. flocculosa.
FUNGAL GENETICS AND BIOLOGY
(2021)
Article
Biochemical Research Methods
Li Liu, Wenbo Li, Ju Li, Dongdong Zhao, Siwei Li, Guo Jiang, Jie Wang, Xuxu Chen, Changhao Bi, Xueli Zhang
Summary: A more stable guide RNA (gRNA), called circular gRNA (cgRNA), was created using a biological technique. Compared to linear gRNA, cgRNA showed increased stability under all tested temperature conditions and maintained its function for 24 hours. By inserting a homologous arm, the circularization efficiency of cgRNA reached 88.2%, and the half-life of the resulting circularized cgRNA was similar to that of purified cgRNA.
ACS SYNTHETIC BIOLOGY
(2023)
Review
Pharmacology & Pharmacy
Hanan Bloomer, Jennifer Khirallah, Yamin Li, Qiaobing Xu
Summary: The CRISPR/Cas system has revolutionized the ability to edit the mammalian genome, providing a platform for correcting pathogenic mutations and studying gene function. Cas9 RNPs, as a delivery method, offer advantages in editing efficiency, off-target editing activity, and cellular toxicity reduction.
ADVANCED DRUG DELIVERY REVIEWS
(2022)
Review
Medicine, Research & Experimental
Song Zhang, Jiangtao Shen, Dali Li, Yiyun Cheng
Summary: CRISPR/Cas9 genome editing as a direct delivery method of RNP complex has advantages of transient genome editing and reduced off-target effects, but faces challenges in efficient material delivery. Current Cas9 RNP delivery systems, including physical approaches and synthetic carriers, are detailed in this review, along with their mechanisms and beneficial roles in intracellular delivery, highlighting examples of stimulus-responsive and targeted carriers in RNP delivery. Challenges of current Cas9 RNP delivery systems and perspectives in rational design of next generation materials for this promising field are discussed.
Article
Agronomy
Sagar Krushnaji Rangari, Manjot Kaur Sudha, Harjot Kaur, Nidhi Uppal, Gagandeep Singh, Yogesh Vikal, Priti Sharma
Summary: This study focused on generating mutants for the ZmPLA1 gene in maize inbred line LM13 through site-directed mutagenesis using CRISPR/Cas9-mediated ribonucleoprotein (RNP) complex method to increase haploid induction rate.
GM CROPS & FOOD-BIOTECHNOLOGY IN AGRICULTURE AND THE FOOD CHAIN
(2023)
Article
Biochemistry & Molecular Biology
Joanna E. Kowalczyk, Shreya Saha, Miia R. Makela
Summary: Dichomitus squalens is an emerging reference species for investigating plant biomass degradation. Recent genomic studies have provided detailed knowledge of the genes and enzymes involved in lignocellulose breakdown in this fungus. Developing efficient genetic manipulation tools is crucial for utilizing this data, as precise genome alterations are often challenging in wild-type basidiomycetes.
Article
Biochemical Research Methods
Stefan J. Tekel, Nicholas Brookhouser, Kylie Standage-Beier, Xiao Wang, David A. Brafman
Summary: The introduction of transient reporters of editing enrichment (TREE) has enabled highly efficient single-base editing of human cells using a transient episomal fluorescent reporter, allowing for rapid generation of clonal editing efficiencies exceeding 80% in biallelic or multiplexed edited isogenic human pluripotent stem cell lines within approximately 3-4 weeks.
Review
Agriculture, Dairy & Animal Science
Julia Popova, Victoria Bets, Elena Kozhevnikova
Summary: Genome editing has practical applications in farm animals, improving production traits, economic value, and disease resistance. It also has potential in biomedical research and drug production, as well as xenograft donors. Recent advancements in site-specific nucleases and embryological delivery methods have revolutionized transgenesis, providing efficient and reliable tools for genome engineering in agriculture.
Review
Biochemistry & Molecular Biology
Yulin Mu, Chengxiao Zhang, Taihua Li, Feng-Jie Jin, Yun-Ju Sung, Hee-Mock Oh, Hyung-Gwan Lee, Long Jin
Summary: Lactobacillus, important in food production and probiotics, can benefit from CRISPR/Cas9-based genome editing to improve strain efficiency and achieve traceless genome modification.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Review
Cell Biology
Joseph Andrew Whitley, Houjian Cai
Summary: Clustered regularly interspaced palindromic repeats (CRISPR) is a gene editing tool with great therapeutic potential. Extracellular vesicles (EVs) have emerged as a promising therapeutic delivery vehicle for CRISPR RNP encapsulation. Different strategies have been identified to encapsulate CRISPR RNP into EVs based on genetic fusion, protein tethering, and sgRNA-coupled encapsulation. Incorporating targeting moieties to EVs membrane surface can increase delivery efficiency. However, there are still limitations preventing the use of these strategies in clinical trials.
JOURNAL OF EXTRACELLULAR VESICLES
(2023)
Article
Agronomy
Catherine Mermans, Wannes Dermauw, Sven Geibel, Thomas Van Leeuwen
Summary: Afoxolaner demonstrated strong antagonistic activity against GABACls of T. urticae, making it a powerful tool for controlling spider mites. In contrast, other drugs such as fluralaner, fipronil, and endosulfan showed minimal antagonistic activities against GABACls channels of T. urticae.
PEST MANAGEMENT SCIENCE
(2023)
Article
Virology
Sofia Balaska, Mattia Calzolari, Annalisa Grisendi, Mara Scremin, Michele Dottori, Konstantinos Mavridis, Romeo Bellini, John Vontas
Summary: The expanding distribution of sand flies, especially in Italy, is leading to an increase in the incidence of visceral leishmaniasis. Regular monitoring is needed to ensure the sustainability and effectiveness of vector control interventions, despite the widespread use of insecticides.
Article
Evolutionary Biology
Ernesto Villacis-Perez, Wenxin Xue, Marilou Vandenhole, Berdien De Beer, Wannes Dermauw, Thomas Van Leeuwen
Summary: Pesticide resistance in the cosmopolitan pest Tetranychus urticae involves various mechanisms, including target-site insensitivity, metabolic detoxification, and additive or synergistic effects. In this study, researchers used experimental evolution and bulked segregant analysis to map quantitative trait loci (QTL) associated with abamectin resistance in two populations of T. urticae. They found shared QTLs containing genes encoding subunits of the glutamate-gated chloride channel (GluCl), as well as novel candidate loci. The study also revealed a non-functional variant of GluCl2 as a unique resistance mechanism. This research uncovers the complex basis of abamectin resistance and highlights the diversity of genetic mechanisms underlying resistance in this pest.
EVOLUTIONARY APPLICATIONS
(2023)
Article
Biochemistry & Molecular Biology
Adam Pym, John G. M. Mina, Bartlomiej J. Troczka, Angela Hayward, Eve Daum, Jan Elias, Russell Slater, John Vontas, Chris Bass, Christoph T. Zimmer
Summary: The tobacco whitefly, Bemisia tabaci, is a polyphagous crop pest that causes significant economic damage worldwide. Neonicotinoids are commonly used insecticides for the control of this species. Understanding the mechanisms responsible for resistance to neonicotinoids in B. tabaci is crucial for effective control and damage limitation. This study reveals that qualitative changes in the cytochrome P450 gene CYP6CM1 alter its metabolic capacity and enhance detoxification of neonicotinoids.
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Wen-Xin Xue, Jing-Tao Sun, Johan Witters, Marilou Vandenhole, Wannes Dermauw, Sabina A. Bajda, Eba A. Simma, Nicky Wybouw, Ernesto Villacis-Perez, Thomas Van Leeuwen
Summary: The two-spotted spider mite Tetranychus urticae has evolved resistance to most classes of pesticides, but different populations show genetic differentiation and reproductive compatibility variation. Understanding these factors is crucial for developing sustainable crop management strategies.
Article
Biochemistry & Molecular Biology
Xueping Lu, Marilou Vandenhole, Dimitra Tsakireli, Spiros A. Pergantis, John Vontas, Wim Jonckheere, Thomas Van Leeuwen
Summary: Acequinocyl and bifenazate, as powerful acaricides, have been found to induce resistance development in spider mites. Target-site resistance resulting from mutations in the Qo pocket of cytochrome b has been identified as the main resistance mechanism. A new mutation, L258F, has been discovered in field populations of Tetranychus urticae, and its role in resistance has been confirmed. However, the resistance in this strain is likely due to a combination of target-site resistance and increased detoxification through P450 enzymes.
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
(2023)
Article
Infectious Diseases
Frederic Labbe, Maha Abdeladhim, Jenica Abrudan, Alejandra Saori Araki, Ricardo N. Araujo, Peter Arensburger, Joshua B. Benoit, Reginaldo Pecanha Brazil, Rafaela V. Bruno, Gustavo Bueno da Silva Rivas, Vinicius Carvalho de Abreu, Jason Charamis, Iliano V. Coutinho-Abreu, Samara G. da Costa-Latge, Alistair Darby, Viv M. Dillon, Scott J. Emrich, Daniela Fernandez-Medina, Nelder Figueiredo Gontijo, Catherine M. Flanley, Derek Gatherer, Fernando A. Genta, Sandra Gesing, Gloria I. Giraldo-Calderon, Bruno Gomes, Eric Roberto Guimaraes Rocha Aguiar, James G. C. Hamilton, Omar Hamarsheh, Mallory Hawksworth, Jacob M. Hendershot, Paul V. Hickner, Jean-Luc Imler, Panagiotis Ioannidis, Emily C. Jennings, Shaden Kamhawi, Charikleia Karageorgiou, Ryan C. Kennedy, Andreas Krueger, Jose M. Latorre-Estivalis, Petros Ligoxygakis, Antonio Carlos A. Meireles-Filho, Patrick Minx, Jose Carlos Miranda, Michael J. Montague, Ronald J. Nowling, Fabiano Oliveira, Joao Ortigao-Farias, Marcio G. Pavan, Marcos Horacio Pereira, Andre Nobrega Pitaluga, Roenick Proveti Olmo, Marcelo Ramalho-Ortigao, Jose M. C. Ribeiro, Andrew J. Rosendale, Mauricio R. V. Sant'Anna, Steven E. Scherer, Nagila F. C. Secundino, Douglas A. Shoue, Caroline da Silva Moraes, Joao Silveira Moledo Gesto, Nataly Araujo Souza, Zainulabueddin Syed, Samuel Tadros, Rayane Teles-de-Freitas, Erich L. Telleria, Chad Tomlinson, Yara M. Traub-Cseko, Joao Trindade Marques, Zhijian Tu, Maria F. Unger, Jesus Valenzuela, Flavia Ferreira, Karla P. V. de Oliveira, Felipe M. Vigoder, John Vontas, Lihui Wang, Gareth D. Weedall, Elyes Zhioua, Stephen Richards, Wesley C. Warren, Robert M. Waterhouse, Rod J. Dillon, Mary Ann McDowell
Summary: Phlebotomine sand flies are important vectors of human disease, transmitting Leishmania parasites that cause devastating diseases. This study sequenced the genomes of two important sand fly vector species and identified genes involved in disease transmission and population structures. The findings provide a foundation for future efforts to prevent the spread of Leishmania parasites.
PLOS NEGLECTED TROPICAL DISEASES
(2023)
Article
Food Science & Technology
Rafaela Panteleri, Amalia Anthousi, Shane Denecke, Debora Boaventura, Ralf Nauen, John Vontas
Summary: The fall armyworm is a global invasive agricultural pest causing major crop losses. Control strategies rely on insecticides and transgenic crops, but resistance development is a significant issue. Mutations in the SfABCC2 gene have been associated with Bt toxin resistance in the fall armyworm. This study expressed SfABCC2 in Drosophila melanogaster to validate resistance mutations and cross-resistance issues.
Article
Agronomy
Emre Inak, Berke Demirci, Marilou Vandenhole, Gokhan Soylemezoglu, Thomas Van Leeuwen, Umut Toprak
Summary: Understanding the molecular mechanisms of resistance and cross-resistance between different ketoenol acaricides is crucial to manage resistance. P450 monooxygenases and carboxyl/cholinesterases were found to be involved in resistance. Gene expression analysis confirmed the involvement of P450-mediated resistance, while no target-site mutations were detected.
Review
Biochemistry & Molecular Biology
Sander De Rouck, Emre Inak, Wannes Dermauw, Thomas Van Leeuwen
Summary: The subclass of Acari, known as Arachnida, consists of numerous harmful pests, including spider mites, Varroa (a bee parasite), Dermanyssus (a poultry mite), and various tick species, which pose threats to agriculture and animal health. To minimize the damage caused by these pests, acaricides are frequently used in agriculture, leading to the development of resistance. Additionally, beneficial predatory mites used in biological control are also affected by acaricide selection. The use of new genetic and genomic tools has greatly advanced our understanding of the molecular genetic mechanisms of resistance in Acari, particularly in the model species Tetranychus urticae. These tools have enabled the discovery and validation of new resistance mutations in a wider range of species and have stimulated research on the gene regulation mechanisms associated with detoxification and resistance.
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Charles Grant, Kumar Saurabh Singh, Angela Hayward, Benjamin J. Hunt, Bartlomiej J. Troczka, Adam Pym, Seung-Joon Ahn, Bin Zeng, Cong-Fen Gao, Alicia Leroux, Eve Daum, Philip Sueess, Dariane Souza, Jan Elias, Richard H. ffrench-Constant, John Vontas, Emmanouil Roditakis, Pablo Bielza, Christoph T. Zimmer, Chris Bass
Summary: Spanish strains of the tomato leafminer exhibit high levels of resistance to the insecticide chlorantraniliprole, which is caused by the overexpression of a UDP-glycosyltransferase gene. The genomic resources generated in this study provide important insights for further research and development of sustainable management strategies for this pest.
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
(2023)
Article
Parasitology
France-Paraudie A. Kouadio, Nadja C. Wipf, Angele S. Nygble, Behi K. Fodjo, Christabelle G. Sadia, John Vontas, Konstantinos Mavridis, Pie Mueller, Chouaibou S. Mouhamadou
Summary: This study aims to investigate the impact of different agricultural practices on phenotypic insecticide resistance and associated molecular resistance mechanisms in Anopheles gambiae s.l. The results show that mosquito populations from all agricultural sites were resistant to several insecticides, with variation in the frequency of resistance mutations and gene expression levels. The findings suggest that agricultural practices might have a role in shaping insecticide resistance profiles in malaria vectors.
PARASITES & VECTORS
(2023)
Article
Agronomy
Christine Njiru, Corinna Saalwaechter, Konstantinos Mavridis, John Vontas, Sven Geibel, Nicky Wybouw, Thomas Van Leeuwen
Summary: A target site mutation in a spider mite pest has been found to confer resistance to certain acaricides and has significant fitness costs. These findings highlight the importance of considering multiple factors in assessing the impact of resistance mutations on pest populations.
PEST MANAGEMENT SCIENCE
(2023)