Efficient collection of a large number of mutations by mutagenesis of DNA damage response defective animals
| Autor: | Katsufumi Dejima, Satoru Iwata, Yuji Suehiro, Sawako Yoshina, Shohei Mitani, Tomoko Motohashi |
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| Rok vydání: | 2021 |
| Předmět: |
DNA Repair
DNA damage Science ved/biology.organism_classification_rank.species Mutagenesis (molecular biology technique) Ataxia Telangiectasia Mutated Proteins Computational biology Gene mutation Biology medicine.disease_cause Genome Article medicine Animals Caenorhabditis elegans Caenorhabditis elegans Proteins Model organism Gene Gene Library Mutation Multidisciplinary Massive parallel sequencing Base Sequence ved/biology High-Throughput Nucleotide Sequencing DNA Sequence Analysis DNA Computer science Phenotype Genetic Techniques Mutagenesis Mutagenesis Site-Directed Medicine DNA Damage Mutagens |
| Zdroj: | Scientific Reports, Vol 11, Iss 1, Pp 1-13 (2021) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | With the development of massive parallel sequencing technology, it has become easier to establish new model organisms that are ideally suited to the specific biological phenomena of interest. Considering the history of research using classical model organisms, we believe that the efficient construction and sharing of gene mutation libraries will facilitate the progress of studies using these new model organisms. Using C. elegans, we applied the TMP/UV mutagenesis method to animals lacking function in the DNA damage response genes atm-1 and xpc-1. This method produces genetic mutations three times more efficiently than mutagenesis of wild-type animals. Furthermore, we confirmed that the use of next-generation sequencing and the elimination of false positives through machine learning could automate the process of mutation identification with an accuracy of over 95%. Eventually, we sequenced the whole genomes of 488 strains and isolated 981 novel mutations generated by the present method; these strains have been made available to anyone who wants to use them. Since the targeted DNA damage response genes are well conserved and the mutagens used in this study are also effective in a variety of species, we believe that our method is generally applicable to a wide range of animal species. |
| Databáze: | OpenAIRE |
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