Giardia duodenalis carries out canonical homologous recombination and single-strand annealing |
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Institution: | 1. Department of Biotechnology and Bioinformatics, Korea University, Sejong 30019, Republic of Korea;2. Interdisciplinary Graduate Program for Artificial Intelligence Smart Convergence Technology, Korea University, Sejong 30019, Republic of Korea;3. Department of Korean Medicine, Semyung University, Jecheon, Chungbuk 27136, Republic of Korea;1. Nazarbayev University, 53 Kabanbay batyr avenue, 010000 Nur-Sultan, Kazakhstan;2. UMR 8126, Institut Gustave Roussy, 114 rue Edouard Vaillant, 94805 Villejuif, France;1. College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China;2. Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China;1. Normandie Univ, UNICAEN U2RM-Stress and Virulence, Esplanade de la Paix, 14032 Caen, France;2. Normandie Univ, LMSM EA4312-Microbiology Signals and Microenvironment, 27000, Evreux, France |
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Abstract: | In the past decades, the ability of Giardia duodenalis to perform homologous recombination has been suggested, supported by the observations of genomic integration of foreign plasmids and the disruption of genes using CRISPR technology. Unfortunately, the direct study of a HR mechanism has not been addressed, which would be pertinent in a minimalist organism lacking fundamental DNA-repair elements and even complete pathways. In addition, the constant ploidy changes through the life cycle of this parasite highlight the conservation and relevance of homologous recombination in maintaining genomic stability. In this research, we analyzed different recombinable plasmid systems and their outcomes after G. duodenalis transfection, using this approach we determined genomic, intra-plasmid and inter-plasmid recombination, moreover, we examined the presence of the non-conservative single-strand annealing pathway. With the intention of corroborating that the observed processes were done by homologous recombination, we used a chemical inhibitor named Mirin, which specifically inhibits Mre11 3′- 5′ exonuclease activity, one of the first steps involved in homologous recombination and fundamental to success in repairing. Overall, these results describe the multiple recombinational substrates used by G. duodenalis to achieve HR and demonstrate the presence and use of single-strand annealing recombination. |
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Keywords: | DNA repair Homologous recombination Mre11 Single-strand annealing DSBs"} {"#name":"keyword" "$":{"id":"kwrd0040"} "$$":[{"#name":"text" "_":"double-strand breaks HR"} {"#name":"keyword" "$":{"id":"kwrd0050"} "$$":[{"#name":"text" "_":"homologous recombination SSA"} {"#name":"keyword" "$":{"id":"kwrd0060"} "$$":[{"#name":"text" "_":"single-strand annealing HA"} {"#name":"keyword" "$":{"id":"kwrd0070"} "$$":[{"#name":"text" "_":"hemagglutinin NHEJ"} {"#name":"keyword" "$":{"id":"kwrd0080"} "$$":[{"#name":"text" "_":"non-homologous end-joining GFP"} {"#name":"keyword" "$":{"id":"kwrd0090"} "$$":[{"#name":"text" "_":"green fluorescent protein |
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