Whole genome sequencing analysis of small and large colony mutants from the mouse lymphoma assay

Guo, Xiao; Pan, Bohu; Seo, Jeong-Woo; Chen, Ying; Yan, Jun; Mei, Nan; Chen, Tao

doi:10.1007/s00204-018-2318-5

Cited by 5 publications

(2 citation statements)

References 40 publications

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“…We anticipate, however, that the newer error-corrected next generation sequencing (EC-NGS) technologies that directly measure mutations at any location in the genome or over the entire genome (Dong et al, 2017;Revollo et al, 2018;Guo et al 2018;Zhang et al, 2019;see Salk, this issue) will: 1) provide a more comprehensive evaluation of smaller gene mutations and hence more appropriate estimates of mutation frequency; and, 2) be able to evaluate gene mutation in any tissue of any animal without the need for using transgene reporters. With the use of longer-read NGS technologies, EC-NGS also may be able to evaluate chromosomal mutation.…”

Section: Recent Developments Issues and Possible Advancementsmentioning

confidence: 99%

Mutation as a Toxicological Endpoint for Regulatory Decision‐Making

Heflich

Johnson

Zeller

et al. 2019

Environ and Mol Mutagen

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Mutations induced in somatic cells and germ cells are responsible for a variety of human diseases, and mutation per se has been considered an adverse health concern since the early part of the 20 th Century. Although in vitro and in vivo somatic cell mutation data are most commonly used by regulatory agencies for hazard identification, i.e., determining whether or not a substance is a potential mutagen and carcinogen, quantitative mutagenicity doseresponse data are being used increasingly for risk assessments. Efforts are currently underway to both improve the measurement of mutations and to refine the computational methods used for evaluating mutation data. We recommend continuing the development of these approaches with the objective of establishing consensus regarding the value of including the quantitative analysis of mutation per se as a required endpoint for comprehensive assessments of toxicological risk.

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Section: Recent Developments Issues and Possible Advancementsmentioning

confidence: 99%

Mutation as a Toxicological Endpoint for Regulatory Decision‐Making

Heflich

Johnson

Zeller

et al. 2019

Environ and Mol Mutagen

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“…Slow growing mutants of both cell types have acquired genetic damage that involves putative growth-regulating gene(s) near the TK locus, resulting in prolonged doubling times and the formation of late appearing or small colonies. More recent studies have demonstrated the molecular nature of mutations in both assays ( Hakulinen et al, 2011 ; Guo et al, 2018 ). Other results indicate that the assays are sensitive enough to detect mutagenicity of NM ( Mei et al, 2012 ; Elespuru et al, 2018 ; Demir et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Thymidine Kinase+/− Mammalian Cell Mutagenicity Assays for Assessment of Nanomaterials

2022

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The methods outlined here are part of a series of papers designed specifically for genotoxicity assessment of nanomaterials (NM). Common Considerations such as NM characterization, sample preparation and dose selection, relevant to all genotoxicity assays, are found in an accompanying paper. The present paper describes methods for evaluation of mutagenicity in the mammalian (mouse) thymidine kinase (Tk) gene occurring in L5178Y mouse lymphoma (ML) cells and in the designated TK gene in human lymphoblastoid TK6 cells. Mutations change the functional genotype from TK+/− to TK−/−, detectable as cells surviving on media selective for the lack of thymidine kinase (TK) function. Unlike cells with TK enzyme function, the TK−/− cells are unable to integrate the toxic selection agent, allowing these cells to survive as rare mutant colonies. The ML assay has been shown to detect a broad spectrum of genetic damage, including both small scale (point) mutations and chromosomal alterations. This assay is a widely used mammalian cell gene mutation assay for regulatory purposes and is included in the core battery of genotoxicity tests for regulatory decision-making. The TK6 assay is an assay using a human cell line derived similarly via mutagenic manipulations and optimal selection. Details are provided on the materials required, cell culture methods, selection of test chemical concentrations, cytotoxicity, treatment time, mutation expression, cloning, and data calculation and interpretation. The methods describe the microwell plate version of the assays without metabolic activation.

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