Report
Targeted insertion of reporter transgene into a gene safe harbor in human blood fluke, Schistosoma mansoni
| العنوان: | Targeted insertion of reporter transgene into a gene safe harbor in human blood fluke, Schistosoma mansoni |
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| المؤلفون: | Tanno, Wannaporn Ittiprasert, Möscheid, Max, F, Chaparro, Cristian, Mann, Victoria, H, Quack, Thomas, Rodpai, Rutchanee, Miller, Andre, Wistiphongpun, Prapakorn, Buakaew, Watunyoo, Mentink-Kane, Margaret, Miller, Legendré, Schmid, Sarah, Popratiloff, Anastas, Grevelding, Christoph, G, Grunau, Christoph, Brindley, Paul, J |
| المساهمون: | The George Washington University (GW), Justus-Liebig-Universität Gießen = Justus Liebig University (JLU), Interactions Hôtes-Pathogènes-Environnements (IHPE), Université de Perpignan Via Domitia (UPVD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Department of Anatomy and Regenerative Biology |
| المصدر: | https://hal.science/hal-03771118 ; 2022. |
| بيانات النشر: | HAL CCSD |
| سنة النشر: | 2022 |
| المجموعة: | Université de Perpignan: HAL |
| مصطلحات موضوعية: | [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology, [SDV.BIO]Life Sciences [q-bio]/Biotechnology |
| الوصف: | We identified genomic safe harbor sites (GSH) in the human blood fluke, Schistosoma mansoni and developed a CRISPR-focused protocol for insertion of a reporter transgene into a representative GSH. The protocol employed ribonuclear protein complexes of Cas9 nuclease, three overlapping guide RNAs, and phosphorothioate-modified, double stranded donor DNAs encoding green fluorescent protein driven by a strong endogenous promoter. Gene-editing efficiencies of >20% and reporter transgene fluorescence of >50% of gene-edited eggs were obtained by five days after CRISPR transfection. These methods and results advance functional genomics for multicellular parasites, and represent a tractable path towards transgenic schistosomes using homology directed repair catalyzed transgene insertion. Identification and characterization of GSH is expected to facilitate consistent transgene activity with neutral influence on the host cell genome and, concurrently, provide a privileged locus for transgene activity. This approach should be adaptable to platyhelminths generally. |
| نوع الوثيقة: | report |
| اللغة: | English |
| Relation: | BIORXIV: 2022.09.02.506379 |
| DOI: | 10.1101/2022.09.02.506379 |
| الاتاحة: | https://hal.science/hal-03771118 https://hal.science/hal-03771118v1/document https://hal.science/hal-03771118v1/file/Ittiprasert-2022-Biorxiv-Targeted-MANUSCRIT.pdf https://doi.org/10.1101/2022.09.02.506379 |
| Rights: | info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.8083657F |
| قاعدة البيانات: | BASE |
| DOI: | 10.1101/2022.09.02.506379 |
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