genetic screens in mice for genome integrity maintenance and cancer predisposition

Genetic screens in mice for genome integrity maintenance andcancer predispositionGabriel Balmus1,2 and Rebecca E McIntyre2

Available online at www.sciencedirect.com

ScienceDirect

Genome instability is a feature of nearly all cancers and can be

exploited for therapy. In addition, a growing number of genome

maintenance genes have been associated with developmental

disorders. Efforts to understand the role of genome instability in

these processes will be greatly facilitated by a more

comprehensive understanding of their genetic network. We

highlight recent genetic screens in model organisms that have

assisted in the discovery of novel regulators of genome stability

and focus on the contribution of mice as a model organism to

understanding the role of genome instability during embryonic

development, tumour formation and cancer therapy.

Addresses1 The Wellcome Trust/Cancer Research UK Gurdon Institute, University

of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK2 Experimental Cancer Genetics, The Wellcome Trust Sanger Institute,

Genome Campus, Hinxton, Cambridge CB10 1SA, UK

Corresponding author: McIntyre, Rebecca E ([email protected])

Current Opinion in Genetics & Development 2014, 24:1–7

This review comes from a themed issue on Cancer genomics

Edited by David J Adams and Ultan McDermott

For a complete overview see the Issue and the Editorial

Available online 13th December 2013

0959-437X/$ – see front matter, # 2013 Elsevier Ltd. All rights

reserved.

http://dx.doi.org/10.1016/j.gde.2013.10.010

IntroductionCells have evolved multiple, elaborate mechanisms to

ensure genome integrity, at the heart of which resides a

complex network of signaling pathways globally known as

the DNA damage response (DDR). The DDR coordi-

nately senses DNA damage, halts the cell cycle and

promotes DNA repair. If repair is not possible, cellular

senescence or apoptosis is activated to ensure that the

defects are not inherited by daughter cells [1,2]. Genome

instability ranges in complexity from point mutations,

gross chromosomal rearrangements or aneuploidy to com-

plete chromosome shattering [3,4].

Genome stability is compromised in many heritable

developmental disorders, including Li-Fraumeni, ataxia

telengiecstasia, xeroderma pigmentosum, Cockayne syn-

drome and trichothiodystrophy (for reviews see [4–7]).

The latter three disorders are all characterised by skin

photosensitivity, but patients with xeroderma pigmento-

sum are highly cancer prone and have an average life

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expectancy of 20 years and those with trichothiodystro-

phy and Cockayne syndrome are not cancer prone but do

age prematurely [8–10]. Despite all three disorders arising

from the inability of the nucleotide excision repair path-

way to repair DNA damage caused by ultraviolet light, we

do not fully understand the differential predisposition to

cancer between these disorders. Moreover, the majority

of spontaneously arising cancer cells are genomically

unstable, but whether this is a consequence of tumour

progression or an active process that drives tumour evol-

ution (the mutator hypothesis) is a question that has still

not been entirely resolved [6,11]. Furthermore, in cancers

with somatically acquired genome instability, it is

possible to manipulate the processes that regulate gen-

ome stability using novel therapies, which results in the

selective killing of tumour cells through catastrophic

genomic instability [12]. Several of these agents are

already showing promise, although cancers may acquire

additional molecular alterations in DNA repair pathways

leading to therapeutic resistance [6,13]. Thus there are

clear disease pathogenesis-driven and therapy-driven

requirements for us to build a more comprehensive net-

work of the processes that regulate genome maintenance

and to understand the interactions between them.

The genes and pathways required to maintain genome

integrity are highly conserved between species and so

mutation of these genes in model organisms can greatly

facilitate both our understanding of human diseases such

as cancer, and the design of therapeutics to improve

health and extend lifespan. Furthermore, the identifi-

cation of novel genome instability genes that suppress

tumourigenesis is of considerable importance for cancer

surveillance programmes and patient care. This review

highlights recent studies of model organisms, especially

mice, that have aided the identification of specific DNA

repair pathways, chromosome transmission and cell cycle

control processes with important consequences for gen-

ome stability, developmental disorders, tumourigenesis

and cancer therapy.

Screening model organisms for genomeinstabilityScreening for genes that maintain genome integrity in

genetically tractable model organisms such as yeast and

mice has considerable advantages over in vitro screens.

Human cancer cell lines such as osteosarcoma-derived

U2OS cells are normally used for in vitro screens [14,15],

but cancer cells carry an array of selectively advantageous

genetic and epigenetic modifications that allow them to


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2 Cancer genomics

Table 1

Mouse models of genomic instability and related human diseases. The table lists genomically unstable mouse mutants that have been

identified through large-scale forward-genetic and reverse-genetic screens. Mice were screened for micronucleus (MN) induction as a

marker of genomic instability [27��,67]. Micronuclei may be fragments of DNA or whole chromosomes and elevated levels of micronuclei

can reflect defects in DNA repair, chromosome transmission or cell-cycle progression. Genomically unstable mutant mice display a wide

spectrum of phenotypes that can inform on the pathogenetic mechanism of human disorders such as cancer

Genetically modified allele MN

Fold-change

Mechanism of genome instability Disease model and comments

Aldh2tm1a(EUCOMM)Wtsi/Hmgu 1.8 Aldehyde dehydrogenase 2 is a mitochondrial protein

required for catabolism of acetaldehydes that damage

DNA.

ALDH2-deficiency and cancer

predisposition [58,68].

Cenpjtm1a(EUCOMM)Wtsi 1.5 Centromere protein J is a centrosomal protein required for

proper chromosome transmission to daughter cells.

Seckel Syndrome. Mice and

patients are not predisposed to

cancer [37�,38].

Mcm4Chaos3 19 Mini-chromosome maintenance deficient 4 homolog (S.

cerevisiae) is part of a complex required for the initiation of

DNA replication.

Familial glucocorticoid deficiency.

Mutant mice are predisposed to

cancer but patient predisposition is

not yet known [28,45,46�,47].

Mcm4D573H 15

Mcph1tm1a(EUCOMM)Wtsi 5 Microcephalin 1 regulates the DNA damage responsive S-

phase and G2/M-phase cell cycle checkpoint and

represses transcription of the human telomerase reverse

transcriptase gene [69,70].

Primary microcephaly

(Mcph1tm1.1Zqw [71]), hearing loss.

Somatic defects, but not germline

defects, are associated with

malignancy [72–74].

Mysm1tm1a(KOMP)Wtsi 2 Myb-like, SWIRM and MPN domains 1-deficiency is

associated with high levels of reactive oxygen species,

which damage DNA, in haematopoietic progenitors [75].

Haematological abnormalities,

human disease not known [75].

Slx4tm1a(EUCOMM)Wtsi 2.5 Structure-specific endonuclease subunit homolog (S.

cerevisiae) is required for DNA double-strand break repair.

Fanconi anaemia cancer

predisposition syndrome [33,34].

Trex1tm1(KOMP)Wtsi 2 30 repair exonuclease 1 is required for nucleic acid

metabolism in the cytosol and nucleus. Clearance of

cytosolic pathogen nucleic acid is part of the innate

immune-response.

Autoimmune diseases, for example

Aicardi-Goutieres syndrome, not

predisposed to malignancy [76,77].

surpass the cellular surveillance apparatus and this ‘back-

ground’ can complicate the interpretation of results. Invitro screens that assess loss-of-function using small inter-

fering RNAs (siRNAs) or short-hairpin RNAs (shRNAs)

may also be complicated by non-specific effects, both at

the mRNA and protein level, and by variable penetrance

of phenotypes due to transfection artifacts. Although

better design of shRNA and siRNAs in recent years

has reduced off-target effects [16,17], there are still issues

with several widely used libraries raising a cautionary note

for such screens. For example, in a recent screen for

regulators of homologous recombination, off-target

depletion of RAD51 was shown to be a common source

of false positives with the Dharmacon human siGEN-

OME siRNA library [15]. The limitations of in vitroscreens highlight the need for in vivo validation of genes

that regulate genome stability in model organisms such as

yeast, zebrafish and mice.

The genotoxicity of environmental and chemical factors

has been monitored in organisms as diverse as bacteria,

zebra mussels, non-human primates and even in humans

following accidental radiation exposure [18–22]. Using

similar methods of detection, large-scale, unbiased

screens of genetically modified model organisms, typi-

cally yeast, have revealed many novel genes that regulate


the cellular responses to different types of exogenous and

endogenous genotoxic assault [23–26,27��]. Disruption of

many of these genes in mice causes genome instability

and several result in phenotypes that are characteristic of

genome instability disorders in humans (Table 1 and

Figure 1). In addition, it is possible to test whether

genetically modified mice are spontaneously tumour

prone or whether a genotoxic insult such as irradiation

can accelerate tumourigenesis (see Table 1) [28]. These

types of genetic screens have been greatly facilitated by

mutation resources for yeast, and more recently for zebra-

fish and mice [23–25,29,30].

Bioinformatics approaches can be useful for increasing

the sensitivity of genome-wide functional genomics

screens by combining datasets and searching for similar

phenotypes. Web-based tools such as Phenomeblast

(http://code.google.com/p/phenomeblast/) enable the

integration, analysis and exploration of phenotypes across

species. Similarly, data from genome-wide screens of the

same species can also be combined to increase power. For

example, data from screens for gross chromosomal re-

arrangements in yeast were combined to search for shared

sensitivity to DNA damaging agents or shared interaction

networks [31�]. The analyses revealed unanticipated roles

for several genes, including a role for the proteasomal


http://code.google.com/p/phenomeblast/

Genetic screens in mice for genome stability Balmus and McIntyre 3

Figure 1

+shRNA+siRNA+cDNA

Human cell lines

Genetic screens for regulators of genome stability

Comparative phenomics

GM Yeast

GM Zebrafish

GM Mice

Neural stem and progenitor cells: microcephaly, intellectual impairment, neurodegeneration.

Haematopoietic stem cells: bone marrow failure.Myeloid progenitor cells:anaemia, predisposition to acute myelogenous leukaemia. Lymphoid progenitor cells: immune deficiency syndromes, predisposition to lymphoma.

Neural crest cells and osteoblasts: craniofacial and skeletal abnormalities, ossification defects, dwarfism.

Skin stem cells: pigmentation defects, light sensitivity, predisposition to skin cancer.

Cell types that are highly sensitive to developmental genomic instability and commonly affected organs

Germline stem cells and spermatogonia:male infertility.

Current Opinion in Genetics & Development

Comparative phenomics is a powerful way to identify human disease-relevant modifiers of genome stability. While large-scale genetic screens in yeast

or human cell lines have been used to identify regulators of genome stability, the same approach in multicellular organisms such as zebrafish and mice

can inform on human disease relevance. The stem cells and stem cell progenitors of humans and mice are highly sensitive to genome damage and

misregulation of the processes that control genome stability results in comparable phenotypes in both species, including predisposition to

malignancies. GM: genetically modified.

subunit RPN10 in the suppression of gross chromosome

rearrangements.

Developmentally acquired genome instabilitydisordersDevelopmental defects in genes that are required for

genome maintenance result in the accumulation of

genetic errors and activation of senescence or apoptosis

in otherwise healthy cells. As with many developmental

diseases, the severity of disease that arises is not only

gene-dependent, but is altered by the precise mutation,

the time point at which the mutation arose during de-

velopment (mosaicism), and the sensitivity of different

stem and progenitor cell types to the defect (Figure 1 and

Table 1). The clinical manifestation of developmental

genome instability disorders is therefore very hetero-

geneous [7]. While genetic screens in yeast and human

cell lines have revealed a role for individual genes in

fundamental biological processes, unbiased large-scale

phenotyping for genomic instability in zebrafish and mice

can help us to better understand the etiology of extremely

rare genetic subtypes of human diseases and may help

clarify the relationship between the genotype and phe-

notype of patients (Figure 1). For example, high-through-

put screens for regulators of genome stability in

genetically modified yeast revealed a role for SLX4 in

homologous recombination [32] and high-throughput


screens for micronucleus formation (a proven marker of

genomic instability and biomarker of cancer predisposi-

tion) in the erythrocytes of loss-of-function mouse

mutants confirmed that Slx4-deficiency causes genome

instability in vivo (Table 1) [33]. Moreover, unbiased

phenotyping of Slx4-deficient mice revealed that they

were infertile and had haematological cytopenias, akin to

patients with the SLX4-deficient genetic subtype of the

cancer predisposition syndrome Fanconi anaemia [33–35].

Similarly, unbiased phenotyping of a Cenpj-hypomorphic

mouse revealed an elevated frequency of micronucleus

formation; however, CENPJ (CPAP) is a centrosomal

protein that had not previously been linked to genome

maintenance. Cenpj-hypomorphic mice shared many

characteristics of patients with the primordial dwarfism

disorder Seckel syndrome and of mice carrying a huma-

nised ATR-hypomorphic allele, including intrauterine

growth retardation, microcephaly and craniofacial

abnormalities (Table 1) [36,37�]. Several Seckel loci have

been identified to date, including ATR, and a single case

report associated mutations in CENPJ with Seckel syn-

drome [7,38]. Embryonic defects in the murine homol-

ogues of ATR and CENPJ cause widespread genetic

instability during development resulting in a high fre-

quency of apoptosis throughout mouse embryos. It is


4 Cancer genomics

thought that this decreases the number of viable cells for

proliferation and growth in utero, resulting in a propor-

tionate reduction in body size. The majority of genes

associated with Seckel syndrome, including ATR, impair

activation of the DDR. However, CENPJ is a centroso-

mal protein and deficiency in mice causes genetic

instability through improper chromosome transmission,

or aneuploidy, demonstrating that model organisms can

inform on new mechanisms of disease [37�]. While Seckel

syndrome has not been associated with a higher incidence

of cancer, most likely because Seckel-cells die before

acquiring adaptive mutations, recently a germline

mutation in ATR was observed in an oropharyngeal cancer

syndrome representing an unexpected clinical outcome

of impaired ATR function [39].

Mosaic variegated aneuploidy (MVA) is a rare autosomal-

recessive genomic instability syndrome characterised by

multiple mosaic aneuploidies in somatic cells. The

clinical manifestations of MVA are similar to CENPJ-

Seckel syndrome and include intrauterine growth retar-

dation, microcephaly and mental retardation; however,

MVA patients are predisposed to rhabdomyosarcoma,

Wilm’s tumour and leukaemia [40]. More than half of

the patients have inactivating mutations in the BUB1Bgene that encodes the BUB1 mitotic (spindle) checkpoint

serine/threonine kinase B, which was first identified by a

genetic screen in yeast [41]. Mitotic spindle checkpoint

proteins arrest the metaphase by inhibiting the anaphase-

promoting complex/cyclosome until the sister chromatids

are correctly attached to the spindle to ensure the main-

tenance of correct chromosome transmission [40]. Bub1b+/

� mice are developmentally normal but have defective

spindle checkpoint activation and develop lung and colon

cancers in response to carcinogens [42]. Structural geno-

mic aberrations that lead to activation of oncogenes or

elimination of tumour suppressor genes have been stu-

died extensively [43]. Furthermore, reduced BUB1B

expression is a feature of chronological ageing and over-

expression of Bub1b in mice was recently shown to protect

against aneuploidy and cancer, and to extend healthy

lifespan in mice. These findings suggest that modulation

of BUB1B activity may be a viable prophylactic therapy

against cancer for both BUB1B-MVA patients and healthy

individuals [44��]. Together, studies of Bub1b-MVA and

Cenpj-Seckel mouse models demonstrate that disruption

of genes within the same biological pathway can result in

similar phenotypes, both at the level of the cell and the

whole organism, but this does not necessarily translate to

a similar tumour predisposition, thus highlighting the

importance of testing this directly in model organisms.

Conversely, studies of model organisms that are genomi-

cally unstable and tumour prone can facilitate the dis-

covery of new disease mechanisms and highlight the need

for close cancer surveillance of patients that carry similar

mutations (Figure 1). For example, several groups have


shown that disruption of Mcm4 in mice leads to genome

instability and tumour predisposition (Table 1) [28,45].

MCM4 is part of an MCM2-7 complex that is required for

DNA replication. Mutation of MCM4 was recently found

to be the underlying molecular defect responsible for a

variant of familial glucocorticoid deficiency [46�,47]. No

defects were observed in the formation of the MCM2-7

complex in fibroblasts from patients, but high levels of re-

replication and DNA breakage were observed [47]. In

support, the adrenal cortices of Mcm4Chaos3 mice were

abnormal and found to contain atypical spindle-shaped

cells [46�]. These studies have demonstrated that familial

glucocorticoid deficiency is a new DNA replication dis-

order. While the patients under investigation did not

show any signs of cancer, the mechanism of their disease

and the tumour predisposition of Mcm4-deficient mice

suggest that these patients should be monitored carefully.

Synthetic interactionsSurgery in combination with chemotherapy remains the

most common approaches for cancer treatment to date

[48]. It is proposed that many if not all cancer cells lack at

least one component of the DDR but since different

DNA-repair pathways can overlap in function, cancer

cells can adapt making it difficult to harness these weak-

nesses [1,13,49]. Following the BRCA/PARP synthetic

lethality success, it is now clear that a better understand-

ing of the synthetic interdependency of the different

DDR pathways can lead to better therapies [50,51]. Thus,

multidisciplinary screening approaches are being used to

discover new synthetic lethal interactions in the context

of personalised cancer medicine [52]. Because of the

accompanying background noise of these biochemical

or cellular screens it is important to confirm these findings

in model organisms. Indeed some of these findings have

been confirmed in mouse models for DDR and cancer.

For example synthetic lethal interactions have been

confirmed between Atr and Trp53/Kras [53], Hus1/Atm[54], Mcm4 and Atm [55], Mcm2/7 [56], Nfkb1 and Trp53/Kras [57] and Fancd2/Aldh2 [58] showing that the success-

ful understanding of the redundancy in cancer relies on

modeling in animal models of disease.

Challenges and future directionsOne of the greatest challenges in this field will be to

understand the rules that govern the relationship be-

tween maintenance of genome stability and tumour for-

mation. Bioinformatics approaches that combine data

from large-scale genetic screens for genome instability

in yeast and mice, together with human cancer gene

expression data from The Cancer Genome Atlas studies,

should make it easier to distinguish between driver and

passenger mutations, which is critical for cancer thera-

peutics.

The use of titratable promoter alleles in yeast will facili-

tate the identification of the rate-limiting factors for



specific DDR pathways, which is both useful in the

design of therapeutics and overcomes a major problem

of loss-of-function genetic screens by enabling the

screening of essential genes for their role in genetic

instability and synthetic lethality [25]. Similarly, a simple

way of increasing detection of gene mutations that cause

embryonic lethality due to catastrophic genetic instability

in mouse genetic screens could be to perform immuno-

histochemistry for gH2AX activation in early embryos.

Several consortia such as ‘DECIPHER’ and ‘UK10K’

have been established to uncover the genetic variants

of unknown developmental disorders and disease-causing

variants [65,66]. It is likely that novel regulators of gen-

ome maintenance will be uncovered by these efforts.

CRISPR-mediated ‘one-step’ genome engineering of

conditional alleles in mice enables the screening of genes

essential for viability and also promises to increase both

the speed and accuracy with which we can model human

disease-relevant mutations [59]. For example knockout of

the DDR gene Atr in mice is embryonic lethal, whereas

mice carrying a humanised hypomorphic allele of ATRmodel the genome instability disorder Seckel syndrome

(Table 1) [60]. Similarly, 48 different mutations in Arte-

mis (DCLRE1C), a DNA nuclease that is required for non-

homologous end-joining, have been identified in associ-

ation with inherited combined immunodeficiency syn-

dromes, including missense, splice-site and nonsense

mutations as well as gross exonic deletions [61]. Unlike

inactivating mutations of Artemis, hypomorphic mutations

of Artemis are hypothesised to predispose to lymphoid

malignancy. In support, generation of a hypomorphic

Artemis human disease allele (ArtP70) in mice results in

a different molecular defect and tumour spectrum when

compared to Artemis nullizygote mice [62]. While

unbiased, large-scale phenotyping programmes designed

to systematically screen loss-of-function (typically

‘knockout’) gene mutations in model organisms have

greatly increased our understanding of gene function,

these examples highlight the need for a more targeted

strategy that aims to phenotype human disease-relevant

mutants.

Finally, DNA sequencing of cancer genomes combined

with the requisite bioinformatics analyses can be used to

tease apart the underlying mutational processes at play in

cancers, including defects in DNA maintenance [63].

Applying the same approach to tumours that form in

animal models harbouring cancer relevant driver

mutations or animals exposed to known genotoxins

may give further insight into the DNA maintenance

processes operative in human cancers and indicate new

avenues for therapeutic intervention [64].

In summary, combining technologies that enable rapid

genome engineering of patient-relevant gene mutations

into model organisms with high-throughput genetic


screens will enable us to explore the role of the genome

maintenance network in human diseases, tumour predis-

position and therapeutics with unprecedented speed and

accuracy. Sequencing human cancers and the tumours

that arise in these model organisms, especially mice, will

enable us to disentangle the defects in DNA maintenance

that contribute to cancer formation from those that do not.

AcknowledgementsWe wish to thank Josep Forment and Kate Dry for their comments. R.E.M.is supported by Cancer Research UK (Project Grant C20510/A12401). G.B.is funded by Cancer Research UK (Program Grant C6/A11224).

References and recommended readingPapers of particular interest, published within the period of review,have been highlighted as:

� of special interest

�� of outstanding interest

1. Jackson SP, Bartek J: The DNA-damage response in humanbiology and disease. Nature 2009, 461:1071-1078.

2. Sirbu BM, Cortez D: DNA damage response: three levels of DNArepair regulation. Cold Spring Harb Perspect Biol 2013,5:a012724.

3. Forment JV, Kaidi A, Jackson SP: Chromothripsis and cancer:causes and consequences of chromosome shattering. Nat RevCancer 2012, 12:663-670.

4. Aguilera A, Gomez-Gonzalez B: Genome instability: amechanistic view of its causes and consequences. Nat RevGenet 2008, 9:204-217.

5. Bunting SF, Nussenzweig A: End-joining, translocations andcancer. Nat Rev Cancer 2013, 13:443-454.

6. Curtin NJ: DNA repair dysregulation from cancer driver totherapeutic target. Nat Rev Cancer 2012, 12:801-817.

7. O’Driscoll M: Diseases associated with defective responses toDNA damage. Cold Spring Harb Perspect Biol 2012, 4:a012773.

8. Vermeulen W, Fousteri M: Mammalian transcription-coupledexcision repair. Cold Spring Harb Perspect Biol 2013, 5:a012625.

9. Stefanini M, Botta E, Lanzafame M, Orioli D: Trichothiodystrophy:from basic mechanisms to clinical implications. DNA Repair(Amst) 2010, 9:2-10.

10. Lehmann AR, McGibbon D, Stefanini M: Xerodermapigmentosum. Orphanet J Rare Dis 2011, 6:70.

11. Hanahan D, Weinberg RA: Hallmarks of cancer: the nextgeneration. Cell 2011, 144:646-674.

12. Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA et al.:Targeting the DNA repair defect in BRCA mutant cells as atherapeutic strategy. Nature 2005, 434:917-921.

13. Bouwman P, Jonkers J: The effects of deregulated DNAdamage signalling on cancer chemotherapy response andresistance. Nat Rev Cancer 2012, 12:587-598.

14. Cotta-Ramusino C, McDonald ER 3rd, Hurov K, Sowa ME,Harper JW et al.: A DNA damage response screen identifiesRHINO, a 9-1-1 and TopBP1 interacting protein required forATR signaling. Science 2011, 332:1313-1317.

15. Adamson B, Smogorzewska A, Sigoillot FD, King RW, Elledge SJ:A genome-wide homologous recombination screen identifiesthe RNA-binding protein RBMX as a component of the DNA-damage response. Nat Cell Biol 2012, 14:318-328.

16. Gu S, Jin L, Zhang Y, Huang Y, Zhang F et al.: The loop position ofshRNAs and pre-miRNAs is critical for the accuracy of dicerprocessing in vivo. Cell 2012, 151:900-911.


http://refhub.elsevier.com/S0959-437X(13)00147-0/sbref0005












































6 Cancer genomics

17. Nolte A, Ott K, Rohayem J, Walker T, Schlensak C et al.:Modification of small interfering RNAs to prevent off-targeteffects by the sense strand. New Biotechnol 2013, 30:159-165.

18. Biran A, Yagur-Kroll S, Pedahzur R, Buchinger S, Reifferscheid Get al.: Bacterial genotoxicity bioreporters. Microb Biotechnol2010, 3:412-427.

19. Michel C, Bourgeault A, Gourlay-France C, Palais F, Geffard Aet al.: Seasonal and PAH impact on DNA strand-break levels ingills of transplanted zebra mussels. Ecotoxicol Environ Saf2013, 92:18-26.

20. Mudry MD, Martinez RA, Nieves M, Carballo MA: Biomarkers ofgenotoxicity and genomic instability in a non-human primate,Cebus libidinosus (Cebidae, Platyrrhini), exposed tonitroimidazole derivatives. Mutat Res 2011, 721:108-113.

21. Nakachi K, Hayashi T, Hamatani K, Eguchi H, Kusunoki Y: Sixtyyears of follow-up of Hiroshima and Nagasaki survivors:current progress in molecular epidemiology studies. Mutat Res2008, 659:109-117.

22. Maluf SW, Passos DF, Bacelar A, Speit G, Erdtmann B:Assessment of DNA damage in lymphocytes of workersexposed to X-radiation using the micronucleus test and thecomet assay. Environ Mol Mutagen 2001, 38:311-315.

23. Alver B, Jauert PA, Brosnan L, O’Hehir M, Vandersluis B et al.: Awhole genome screen for minisatellite stability genes instationary phase yeast cells. G3 (Bethesda) 2013, 3:741-756.

24. Cheng E, Vaisica JA, Ou J, Baryshnikova A, Lu Y et al.: Genomerearrangements caused by depletion of essential DNAreplication proteins in Saccharomyces cerevisiae. Genetics2012, 192:147-160.

25. Stirling PC, Bloom MS, Solanki-Patil T, Smith S, Sipahimalani Pet al.: The complete spectrum of yeast chromosome instabilitygenes identifies candidate CIN cancer genes and functionalroles for ASTRA complex components. PLoS Genet 2011,7:e1002057.

26. Choy JS, O’Toole E, Schuster BM, Crisp MJ, Karpova TS et al.:Genome-wide haploinsufficiency screen reveals a novel rolefor gamma-TuSC in spindle organization and genomestability. Mol Biol Cell 2013, 24:2753-2763.

27.��

White JK, Gerdin AK, Karp NA, Ryder E, Buljan M et al.: Genome-wide generation and systematic phenotyping of knockoutmice reveals new roles for many genes. Cell 2013, 154:452-464.

This article describes the high-throughput, unbiased phenotyping of loss-of-function mouse mutants and includes a screen for micronucleusinduction (at 16 weeks of age) as a marker of genomic instability.

28. Shima N, Alcaraz A, Liachko I, Buske TR, Andrews CA et al.: Aviable allele of Mcm4 causes chromosome instability andmammary adenocarcinomas in mice. Nat Genet 2007, 39:93-98.

29. Skarnes WC, Rosen B, West AP, Koutsourakis M, Bushell W et al.:A conditional knockout resource for the genome-wide studyof mouse gene function. Nature 2011, 474:337-342.

30. Kettleborough RN, Busch-Nentwich EM, Harvey SA, Dooley CM,de Bruijn E et al.: A systematic genome-wide analysis ofzebrafish protein-coding gene function. Nature 2013, 496:494-497.

31.�

Putnam CD, Allen-Soltero SR, Martinez SL, Chan JE, Hayes TKet al.: Bioinformatic identification of genes suppressinggenome instability. Proc Natl Acad Sci U S A 2012, 109:E3251-E3259.

This article demonstrates how bioinformatics can increase the sensitivityand power of large-scale genetic screens to find new regulators ofgenome stability.

32. Coulon S, Gaillard PH, Chahwan C, McDonald WH, Yates JR 3rdet al.: Slx1-Slx4 are subunits of a structure-specificendonuclease that maintains ribosomal DNA in fission yeast.Mol Biol Cell 2004, 15:71-80.

33. Crossan GP, van der Weyden L, Rosado IV, Langevin F,Gaillard PH et al.: Disruption of mouse Slx4, a regulator ofstructure-specific nucleases, phenocopies Fanconi anemia.Nat Genet 2011, 43:147-152.


34. Stoepker C, Hain K, Schuster B, Hilhorst-Hofstee Y, Rooimans MAet al.: SLX4, a coordinator of structure-specificendonucleases, is mutated in a new Fanconi anemia subtype.Nat Genet 2011, 43:138-141.

35. Holloway JK, Mohan S, Balmus G, Sun X, Modzelewski A et al.:Mammalian BTBD12 (SLX4) protects against genomicinstability during mammalian spermatogenesis. PLoS Genet2011, 7:e1002094.

36. Murga M, Bunting S, Montana MF, Soria R, Mulero F et al.: Amouse model of ATR-Seckel shows embryonic replicativestress and accelerated aging. Nat Genet 2009, 41:891-898.

37.�

McIntyre RE, Lakshminarasimhan Chavali P, Ismail O,Carragher DM, Sanchez-Andrade G et al.: Disruption of mouseCenpj, a regulator of centriole biogenesis, phenocopiesSeckel syndrome. PLoS Genet 2012, 8:e1003022.

This article demonstrates that mouse models of human diseases can beused to identify subtly different mechanisms for genetic subtypes ofhuman diseases. The majority of cells from Seckel patients have a defectin the DNA damage response that lowers the threshold for apoptosis.Cells from Cenpj-Seckel mice were proficient in their activation of theDNA damage response and instead, showed increased chromosomalinstability, which also lowers the threshold for apoptosis.

38. Al-Dosari MS, Shaheen R, Colak D, Alkuraya FS: Novel CENPJmutation causes Seckel syndrome. J Med Genet 2010, 47:411-414.

39. Tanaka A, Weinel S, Nagy N, O’Driscoll M, Lai-Cheong JE et al.:Germline mutation in ATR in autosomal-dominantoropharyngeal cancer syndrome. Am J Hum Genet 2012,90:511-517.

40. Ganmore I, Smooha G, Izraeli S: Constitutional aneuploidy andcancer predisposition. Hum Mol Genet 2009, 18:R84-R93.

41. Roberts BT, Farr KA, Hoyt MA: The Saccharomyces cerevisiaecheckpoint gene BUB1 encodes a novel protein kinase. MolCell Biol 1994, 14:8282-8291.

42. Dai W, Wang Q, Liu T, Swamy M, Fang Y et al.: Slippage of mitoticarrest and enhanced tumor development in mice with BubR1haploinsufficiency. Cancer Res 2004, 64:440-445.

43. Baker DJ, Jin F, Jeganathan KB, van Deursen JM: Wholechromosome instability caused by Bub1 insufficiency drivestumorigenesis through tumor suppressor gene loss ofheterozygosity. Cancer Cell 2009, 16:475-486.

44.��

Baker DJ, Dawlaty MM, Wijshake T, Jeganathan KB, Malureanu Let al.: Increased expression of BubR1 protects againstaneuploidy and cancer and extends healthy lifespan. Nat CellBiol 2013, 15:96-102.

Reduced BUB1B (BUBR1) expression is a feature of chronologicalaging. This study uses a transgenic approach in mice to demonstratethat overexpression of Bub1b can counteract defects that causewhole-chromosome instability, even in the presence of genetic alterationsthat strongly promote aneuploidy and cancer such as oncogenic Ras.This suggests that modulation of BUB1B activity may extend healthylifespan.

45. Bagley BN, Keane TM, Maklakova VI, Marshall JG, Lester RA et al.:A dominantly acting murine allele of Mcm4 causeschromosomal abnormalities and promotes tumorigenesis.PLoS Genet 2012, 8:e1003034.

46.�

Hughes CR, Guasti L, Meimaridou E, Chuang CH, Schimenti JCet al.: MCM4 mutation causes adrenal failure, short stature,and natural killer cell deficiency in humans. J Clin Invest 2012,122:814-820.

This is an excellent example of how shared phenotypes between geneti-cally modified mice and patients lead to greater certainty in the patho-genic nature of disease. The article demonstrates how previousmechanistic studies of Mcm4 disruption in mice helped to assign anew mechanism of disease for familial glucocorticoid deficiency. Impor-tantly, previous studies showed that Mcm4-deficient mice were tumourprone suggesting that these patients will require careful monitoring forcancer formation.

47. Gineau L, Cognet C, Kara N, Lach FP, Dunne J et al.: PartialMCM4 deficiency in patients with growth retardation, adrenalinsufficiency, and natural killer cell deficiency. J Clin Invest2012, 122:821-832.




















































































































48. Chabner BA, Roberts TG Jr: Timeline: chemotherapy and thewar on cancer. Nat Rev Cancer 2005, 5:65-72.

49. Canaani D: Application of the concept synthetic lethality towardanticancer therapy: a promise fulfilled? Cancer Lett 2013.

50. Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D et al.:Specific killing of BRCA2-deficient tumours with inhibitors ofpoly(ADP-ribose) polymerase. Nature 2005, 434:913-917.

51. Lord CJ, Ashworth A: The DNA damage response and cancertherapy. Nature 2012, 481:287-294.

52. von Stechow L, van de Water B, Danen EH: Unraveling DNAdamage response-signaling networks through systemsapproaches. Arch Toxicol 2013, 87:1635-1648.

53. Gilad O, Nabet BY, Ragland RL, Schoppy DW, Smith KD et al.:Combining ATR suppression with oncogenic Rassynergistically increases genomic instability, causingsynthetic lethality or tumorigenesis in a dosage-dependentmanner. Cancer Res 2010, 70:9693-9702.

54. Balmus G, Zhu M, Mukherjee S, Lyndaker AM, Hume KR et al.:Disease severity in a mouse model of ataxia telangiectasia ismodulated by the DNA damage checkpoint gene Hus1. HumMol Genet 2012, 21:3408-3420.

55. Wallace MD, Southard TL, Schimenti KJ, Schimenti JC: Role ofDNA damage response pathways in preventingcarcinogenesis caused by intrinsic replication stress.Oncogene 2013.

56. Chuang CH, Yang D, Bai G, Freeland A, Pruitt SC et al.: Post-transcriptional homeostasis and regulation of MCM2-7 inmammalian cells. Nucleic Acids Res 2012, 40:4914-4924.

57. Meylan E, Dooley AL, Feldser DM, Shen L, Turk E et al.:Requirement for NF-kappaB signalling in a mouse model oflung adenocarcinoma. Nature 2009, 462:104-107.

58. Langevin F, Crossan GP, Rosado IV, Arends MJ, Patel KJ: Fancd2counteracts the toxic effects of naturally produced aldehydesin mice. Nature 2011, 475:53-58.

59. Yang H, Wang H, Shivalila CS, Cheng AW, Shi L et al.: One-stepgeneration of mice carrying reporter and conditional alleles byCRISPR/Cas-mediated genome engineering. Cell 2013,154:1370-1379.

60. O’Driscoll M: Mouse models for ATR deficiency. DNA Repair(Amst) 2009, 8:1333-1337.

61. Pamela P, Leea LW, Gilmoura KC, Bibic S, Calea CM, Amroliaa PJ,Veysa PA, Graham Daviesa E, Jeggob PA, Jonesa A: The manyfaces of artemis-deficient combined immunodeficiency — twopatients with DCLRE1C mutations and a systematic literaturereview of genotype-phenotype correlation. Clin Immunol 2013.

62. Jacobs C, Huang Y, Masud T, Lu W, Westfield G et al.: Ahypomorphic Artemis human disease allele causes aberrantchromosomal rearrangements and tumorigenesis. Hum MolGenet 2011, 20:806-819.


63. Alexandrov LB, Nik-Zainal S, Wedge DC, Aparicio SA, Behjati Set al.: Signatures of mutational processes in human cancer.Nature 2013, 500:415-421.

64. McIntyre RE, van der Weyden L, Adams DJ: Cancer genediscovery in the mouse. Curr Opin Genet Dev 2012, 22:14-20.

65. Firth HV, Wright CF, Study DDD: The deciphering developmentaldisorders (DDD) study. Dev Med Child Neurol 2011, 53:702-703.

66. UK10K — Rare Genetic Variants in Health and Disease. http://www.uk10k.org/.

67. Reinholdt L, Ashley T, Schimenti J, Shima N: Forward geneticscreens for meiotic and mitotic recombination-defectivemutants in mice. Methods Mol Biol 2004, 262:87-107.

68. Garaycoechea JI, Crossan GP, Langevin F, Daly M, Arends MJet al.: Genotoxic consequences of endogenous aldehydes onmouse haematopoietic stem cell function. Nature 2012,489:571-575.

69. Singh N, Basnet H, Wiltshire TD, Mohammad DH, Thompson JRet al.: Dual recognition of phosphoserine and phosphotyrosinein histone variant H2A.X. by DNA damage response proteinMCPH1. Proc Natl Acad Sci U S A 2012, 109:14381-14386.

70. Shi L, Li M, Su B: MCPH1/BRIT1 represses transcription of thehuman telomerase reverse transcriptase gene. Gene 2012,495:1-9.

71. Gruber R, Zhou Z, Sukchev M, Joerss T, Frappart PO et al.:MCPH1 regulates the neuroprogenitor division mode bycoupling the centrosomal cycle with mitotic entry through theChk1-Cdc25 pathway. Nat Cell Biol 2011, 13:1325-1334.

72. Chen J, Ingham N, Clare S, Raisen C, Vancollie VE et al.: Mcph1-deficient mice reveal a role for MCPH1 in otitis media. PLoSONE 2013, 8:e58156.

73. Jo YH, Kim HO, Lee J, Lee SS, Cho CH et al.: MCPH1 proteinexpression and polymorphisms are associated with risk ofbreast cancer. Gene 2013, 517:184-190.

74. Bhattacharya N, Mukherjee N, Singh RK, Sinha S, Alam N et al.:Frequent alterations of MCPH1 and ATM are associated withprimary breast carcinoma: clinical and prognosticimplications. Ann Surg Oncol 2012.

75. Nijnik A, Clare S, Hale C, Raisen C, McIntyre RE et al.: The criticalrole of histone H2A-deubiquitinase Mysm1 in hematopoiesisand lymphocyte differentiation. Blood 2012, 119:1370-1379.

76. Morita M, Stamp G, Robins P, Dulic A, Rosewell I et al.: Gene-targeted mice lacking the Trex1 (DNase III) 30 ! 50 DNAexonuclease develop inflammatory myocarditis. Mol Cell Biol2004, 24:6719-6727.

77. Crow YJ, Rehwinkel J: Aicardi-Goutieres syndrome and relatedphenotypes: linking nucleic acid metabolism withautoimmunity. Hum Mol Genet 2009, 18:R130-R136.


























































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