New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay
Abstract Background Gene mutation assays in transgenic rodents are useful tools to investigate in vivo mutagenicity in a target tissue. Using a lambda EG10 transgene containing reporter genes, gpt delta transgenic mice and rats have been developed to detect point mutations and deletions. The transge...
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doaj-d36d5fcbf94e4113beef74e2f5e7d3f12021-06-27T11:46:12ZengBMCGenes and Environment1880-70622021-06-0143111210.1186/s41021-021-00195-1New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assayKenichi Masumura0Tomoko Ando1Akiko Ukai2Sho Fujiwara3Shigeo Yokose4Xinyue You5Takayoshi Suzuki6Hiroyuki Hayashi7Takehiko Nohmi8Hisayoshi Takagi9Masamitsu Honma10Division of Genetics and Mutagenesis, National Institute of Health SciencesDivision of Genetics and Mutagenesis, National Institute of Health SciencesDivision of Genetics and Mutagenesis, National Institute of Health SciencesBiotechnical Center, Japan SLC, Inc.Biotechnical Center, Japan SLC, Inc.Division of Molecular Target and Gene Therapy Products, National Institute of Health SciencesSchool of Public Health, Hongqiao International Institute of Medicine, Shanghai Jiao Tong University School of MedicineBiologics Business Planning & Operation Dept, Meiji Seika Pharma Co., LtdDivision of Pathology, National Institute of Health SciencesBiotechnical Center, Japan SLC, Inc.Division of Genetics and Mutagenesis, National Institute of Health SciencesAbstract Background Gene mutation assays in transgenic rodents are useful tools to investigate in vivo mutagenicity in a target tissue. Using a lambda EG10 transgene containing reporter genes, gpt delta transgenic mice and rats have been developed to detect point mutations and deletions. The transgene is integrated in the genome and can be rescued through an in vitro packaging reaction. However, the packaging efficiency is lower in gpt delta rats than in mice, because of the transgene in gpt delta rats being heterozygous and in low copy number. To improve the packaging efficiency, we herein describe a newly developed homozygous gpt delta rat strain. Results The new gpt delta rat has a Wistar Hannover background and has been successfully maintained as homozygous for the transgene. The packaging efficiency in the liver was 4 to 8 times higher than that of existing heterozygous F344 gpt delta rats. The frequency of gpt point mutations significantly increased in the liver and bone marrow of N-nitroso-N-ethylurea (ENU)- and benzo[a]pyrene (BaP)-treated rats. Spi− deletion frequencies significantly increased in the liver and bone marrow of BaP-treated rats but not in ENU-treated rats. Whole genome sequencing analysis identified ≥ 30 copies of lambda EG10 transgenes integrated in rat chromosome 1. Conclusions The new homozygous gpt delta rat strain showed a higher packaging efficiency, and could be useful for in vivo gene mutation assays in rats.https://doi.org/10.1186/s41021-021-00195-1gpt delta transgenic ratgpt assaySpi− assaymutant frequencymutation spectrum |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kenichi Masumura Tomoko Ando Akiko Ukai Sho Fujiwara Shigeo Yokose Xinyue You Takayoshi Suzuki Hiroyuki Hayashi Takehiko Nohmi Hisayoshi Takagi Masamitsu Honma |
spellingShingle |
Kenichi Masumura Tomoko Ando Akiko Ukai Sho Fujiwara Shigeo Yokose Xinyue You Takayoshi Suzuki Hiroyuki Hayashi Takehiko Nohmi Hisayoshi Takagi Masamitsu Honma New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay Genes and Environment gpt delta transgenic rat gpt assay Spi− assay mutant frequency mutation spectrum |
author_facet |
Kenichi Masumura Tomoko Ando Akiko Ukai Sho Fujiwara Shigeo Yokose Xinyue You Takayoshi Suzuki Hiroyuki Hayashi Takehiko Nohmi Hisayoshi Takagi Masamitsu Honma |
author_sort |
Kenichi Masumura |
title |
New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
title_short |
New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
title_full |
New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
title_fullStr |
New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
title_full_unstemmed |
New homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
title_sort |
new homozygous gpt delta transgenic rat strain improves an efficiency of the in vivo mutagenicity assay |
publisher |
BMC |
series |
Genes and Environment |
issn |
1880-7062 |
publishDate |
2021-06-01 |
description |
Abstract Background Gene mutation assays in transgenic rodents are useful tools to investigate in vivo mutagenicity in a target tissue. Using a lambda EG10 transgene containing reporter genes, gpt delta transgenic mice and rats have been developed to detect point mutations and deletions. The transgene is integrated in the genome and can be rescued through an in vitro packaging reaction. However, the packaging efficiency is lower in gpt delta rats than in mice, because of the transgene in gpt delta rats being heterozygous and in low copy number. To improve the packaging efficiency, we herein describe a newly developed homozygous gpt delta rat strain. Results The new gpt delta rat has a Wistar Hannover background and has been successfully maintained as homozygous for the transgene. The packaging efficiency in the liver was 4 to 8 times higher than that of existing heterozygous F344 gpt delta rats. The frequency of gpt point mutations significantly increased in the liver and bone marrow of N-nitroso-N-ethylurea (ENU)- and benzo[a]pyrene (BaP)-treated rats. Spi− deletion frequencies significantly increased in the liver and bone marrow of BaP-treated rats but not in ENU-treated rats. Whole genome sequencing analysis identified ≥ 30 copies of lambda EG10 transgenes integrated in rat chromosome 1. Conclusions The new homozygous gpt delta rat strain showed a higher packaging efficiency, and could be useful for in vivo gene mutation assays in rats. |
topic |
gpt delta transgenic rat gpt assay Spi− assay mutant frequency mutation spectrum |
url |
https://doi.org/10.1186/s41021-021-00195-1 |
work_keys_str_mv |
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