Analysis of Microsatellite Polymorphism in Inbred Knockout Mice

BaoFen, Zuo; Du, Xiaoyan; Zhao, Jing; Yang, Huixin; Wang, Chao; Wu, Yongfeng; Lü, Jing; Wang, Ying; Chen, Zhenwen

doi:10.1371/journal.pone.0034555

Cited by 17 publications

(28 citation statements)

References 34 publications

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“…This may be due to differences in the route and dose, and duration of E2 administration, or the effects of progesterone in mice with ovaries. MNU reportedly evokes not only various types of DNA damage [25][26][27], but also MSI and subsequent point mutations [30][31][32]. A previous study reported that an intraperitoneal injection of MNU into CD mice resulted in the development of thymic lymphoma and systemic MSI [30], suggesting that MNU impairs MMR function.…”

Section: Discussionmentioning

confidence: 99%

Elevated Serum Levels of Estradiol Induce Endometrial Hyperplasia Rather than Carcinoma in a Mouse Model Using a Carcinogen

Asaka¹,

Miyamoto²,

Yumura³

et al. 2017

J Carcinog Mutagen

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Objective: Although estrogen has been regarded as a risk factor for endometrial carcinoma, its concentrationdependent carcinogenetic effects remain unclear, because most endometrial carcinomas occur in post-menopausal women, whose serum estrogen levels are relatively low. We previously reported that high levels of estradiol (E2) may suppress endometrial carcinogenesis by up-regulating DNA mismatch repair (MMR) in vitro. The present study was undertaken to further examine the carcinogenetic role of estrogen at various concentrations in vivo.Methods: N-methyl-N-nitrosourea (MNU) was injected into the uterine cavity of 29 mice, and E2 was administered by pellets or orally. Uteri were removed for histological examinations 24 weeks later, and serum E2 levels were measured. The immunohistochemical expression of MMR proteins in uterine epithelia was investigated.Results: Of 29 mice, 8, 8, 8, and 5 showed atrophic, normal, hyperplastic, and carcinomatous endometria, respectively. The mean E2 levels of each group were 0.2 pg/ml, 3.8 pg/ml, 190.0 pg/ml, and 6.7 pg/ml, with significant differences. The expression of the MMR proteins was stronger in mice with elevated E2. Conclusion:Elevated E2 levels preferentially induced endometrial hyperplasia rather than carcinoma, and this may be mediated by MMR proteins. These results indicate that modest E2 is needed, whereas elevated E2 levels are not necessarily advantageous for carcinogenesis, suggesting the importance of low-chronic (un-opposed) estrogen in human endometrial carcinogenesis.

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Section: Discussionmentioning

confidence: 99%

Elevated Serum Levels of Estradiol Induce Endometrial Hyperplasia Rather than Carcinoma in a Mouse Model Using a Carcinogen

Asaka¹,

Miyamoto²,

Yumura³

et al. 2017

J Carcinog Mutagen

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“…The hybrid scaffolded assembly was polished using Quiver 10 to improve consensus accuracies in the range of Q60 and to reduce the high indel errors that are expected in the PacBio sequencing data 9 . pbalign (https://github.com/PacificBiosciences/pbalign) was used to create a bam file from all h5 files of SMRT cells and Quiver trained on P6-C4 chemistries were used to obtain the consensus corrected assembly.…”

Section: Polishing and Assembly Evaluationmentioning

confidence: 99%

The genome of C57BL/6J “Eve”, the mother of the laboratory mouse genome reference strain

Sarsani

Raghupathy

Fiddes

et al. 2019

Preprint

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Isogenic laboratory mouse strains are used to enhance reproducibility as individuals within a strain are essentially genetically identical. For the most widely used isogenic strain, C57BL/6, there is also a wealth of genetic, phenotypic, and genomic data, including one of the highest quality reference genomes (GRCm38.p6). However, laboratory mouse strains are living reagents and hence genetic drift occurs and is an unavoidable source of accumulating genetic variability that can have an impact on reproducibility over time. Nearly 20 years after the first release of the mouse reference genome, individuals from the strain it represents (C57BL/6J) are at least 26 inbreeding generations removed from the individuals used to generate the mouse reference genome. Moreover, C57BL/6J is now maintained through the periodic reintroduction of mice from cryopreserved embryo stocks that are derived from a single breeder pair, aptly named C57BL/6J Adam and Eve. To more accurately represent the genome of today's C57BL/6J mice, we have generated a de novo assembly of the C57BL/6J Eve genome (B6Eve) using high coverage, long-read sequencing, optical mapping, and short-read data. Using these data, we addressed recurring variants observed in previous mouse studies. We have also identified structural variations that impact coding sequences, closed gaps in the mouse reference assembly, some of which are in genes, and we have identified previously unannotated coding sequences through long read sequencing of cDNAs. This B6Eve assembly explains discrepant observations that have been associated with GRCm38-based analyses, and has provided data towards a reference genome that is more representative of the C57BL/6J mice that are in use today.

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“…Primers were fluorescently labeled with 6-FAM (blue), HEX (green) and TAMRA (yellow), for which the internal size standard was ROX (red). The PCR procedure was carried out as previously described (Zuo et al, 2012). Briefly, primers were first tagged to dyes at the 5' ends and PCR performed using 15 μL volume reactions.…”

Section: Stage Three: Capillary Electrophoresis-based Screeningmentioning

confidence: 99%

Development of novel DNA markers for genetic analysis of grey hamsters by cross-species amplification of microsatellites

Wang

Zhang

et al. 2015

Genet. Mol. Res.

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ABSTRACT. The grey hamster has been used in biomedical research for decades. However, effective molecular methods for evaluating the genetic structure of this species are lacking, which hinders its wider usage. In this study, we employed cross-amplification of microsatellite loci of species within the same genus by polymerase chain reaction. Loci screened included 107 from the Mongolian gerbil (MG) and 60 from the Chinese hamster (CH); of these, 15 polymorphic loci were identified for the grey hamster. Of the 167 loci screened, 95 (56.9%) with clear bands on agarose gel were initially identified. After sequencing, 74 (77.9%) of these matched the criteria for microsatellite characteristics, including 41 from MG and 33 from CH. Lastly, 15 (20.3%) loci with more than two alleles for each locus were identified through capillary electrophoresis scanning. To justify the applicability of the 15 grey hamster loci, genetic indexes of grey hamsters were evaluated using 46 generations of outbred stock, established 20 years ago, from Xinjiang, China. Mean effective allele numbers and expected heterozygosity of stock were as low as, respectively, 1.2 and 0.14; these were 2.8 and 4.0 times inferior, respectively, 14339-14347 (2015) to wild grey hamsters. This finding suggests that the genetic structure of the stock-bred population is too weak to resist artificial and natural selection, mutation and genetic drifting. In conclusion, we have developed de novo microsatellite markers for genetic analysis of the grey hamster, providing data and methodology for the enrichment of a genetic library for this species.

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Analysis of Microsatellite Polymorphism in Inbred Knockout Mice

Cited by 17 publications

References 34 publications

Elevated Serum Levels of Estradiol Induce Endometrial Hyperplasia Rather than Carcinoma in a Mouse Model Using a Carcinogen

Elevated Serum Levels of Estradiol Induce Endometrial Hyperplasia Rather than Carcinoma in a Mouse Model Using a Carcinogen

The genome of C57BL/6J “Eve”, the mother of the laboratory mouse genome reference strain

Development of novel DNA markers for genetic analysis of grey hamsters by cross-species amplification of microsatellites

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